smo_human
NAME
5V57
ORGANISM
Human (Homo sapiens)
PROTEIN NAME
smo_human
SOURCE
SWISSPROT
SEQUENCE
Perspective:
Wild-type
Construct
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N-term | |||||||||
M
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A
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A
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A
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R
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P
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A
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R
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G
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P
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10 |
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E
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L
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P
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L
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L
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G
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L
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L
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L
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L
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20 |
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L
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L
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L
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G
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D
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P
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G
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R
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G
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A
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30 |
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A
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S
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S
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G
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N
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A
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T
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G
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P
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G
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40 |
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P
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R
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S
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A
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G
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G
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S
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A
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R
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R
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50 |
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|||
S
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A
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A
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V
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T
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G
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P
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P
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P
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P
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60 |
L
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S
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H
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C
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G
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R
|
A
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A
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P
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C
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70 |
E
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P
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L
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R
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Y
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N
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V
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C
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L
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G
|
80 |
S
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V
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L
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P
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Y
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G
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A
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T
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S
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T
|
90 |
L
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L
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A
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G
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D
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S
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D
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S
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Q
|
E
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100 |
E
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A
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H
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G
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K
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L
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V
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L
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W
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S
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110 |
G
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L
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R
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N
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A
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P
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R
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C
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W
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A
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120 |
V
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I
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Q
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P
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L
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L
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C
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A
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V
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Y
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130 |
M
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P
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K
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C
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E
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N
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D
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R
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V
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E
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140 |
L
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P
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S
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R
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T
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L
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C
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Q
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A
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T
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150 |
R
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G
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P
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C
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A
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I
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V
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E
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R
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E
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160 |
R
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G
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W
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P
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D
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F
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L
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R
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C
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T
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170 |
P
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D
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R
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F
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P
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E
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G
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C
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T
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N
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180 |
E
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V
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Q
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N
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I
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K
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F
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N
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S
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S
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190 |
G
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Q
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C
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E
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V
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P
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L
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V
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R
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T
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200 |
D
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N
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P
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K
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S
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W
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Y
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E
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D
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V
|
210 |
E
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G
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C
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G
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I
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Q
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C
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Q
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N
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P
|
220 |
TM1 | |||||||||
L
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F
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T
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E
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A
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E
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H
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Q
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D
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M
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230 |
H
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S
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Y
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I
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A
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A
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F
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G
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A
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V
|
240 |
T
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G
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L
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C
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T
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L
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F
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T
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L
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A
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250 |
ICL1 | |||||||||
T
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F
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V
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A
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D
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W
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R
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N
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S
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N
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260 |
TM2 | |||||||||
R
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Y
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P
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A
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V
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I
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L
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F
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Y
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V
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270 |
N
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A
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C
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F
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F
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V
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G
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S
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I
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G
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280 |
ECL1 | |||||||||
W
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L
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A
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Q
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F
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M
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D
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G
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A
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R
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290 |
R
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E
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I
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V
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C
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R
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A
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D
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G
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T
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300 |
M
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R
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L
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G
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E
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P
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T
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S
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N
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E
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310 |
TM3 | |||||||||
T
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L
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S
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C
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V
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I
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I
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F
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V
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I
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320 |
V
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Y
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Y
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A
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L
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M
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A
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G
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V
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V
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330 |
W
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F
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V
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V
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L
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T
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Y
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A
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W
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H
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340 |
ICL2 | |||||||||
T
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S
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F
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K
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A
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L
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G
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T
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T
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Y
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350 |
TM4 | |||||||||
Q
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P
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L
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S
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G
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K
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T
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S
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Y
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F
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360 |
H
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L
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L
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T
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W
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S
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L
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P
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F
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V
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370 |
L
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T
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V
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A
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I
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L
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A
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V
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A
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Q
|
380 |
ECL2 | |||||||||
V
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D
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G
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D
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S
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V
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S
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G
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I
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C
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390 |
TM5 | |||||||||
F
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V
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G
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Y
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K
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N
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Y
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R
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Y
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R
|
400 |
A
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G
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F
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V
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L
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A
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P
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I
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G
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L
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410 |
V
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L
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I
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V
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G
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G
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Y
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F
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L
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I
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420 |
R
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G
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V
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M
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T
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L
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F
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S
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I
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K
|
430 |
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ICL3 | TM6 | ||||||||
S
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N
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H
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P
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G
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L
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L
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S
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E
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K
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440 |
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|||||||
A
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A
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S
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K
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I
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N
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E
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T
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M
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L
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450 |
R
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L
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G
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I
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F
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G
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F
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L
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A
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F
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460 |
G
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F
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V
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L
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I
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T
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F
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S
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C
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H
|
470 |
F
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Y
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D
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F
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F
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N
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Q
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A
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E
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W
|
480 |
E
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R
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S
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F
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R
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D
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Y
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V
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L
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C
|
490 |
ECL3 | |||||||||
Q
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A
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N
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V
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T
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I
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G
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L
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P
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T
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500 |
K
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Q
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P
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I
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P
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D
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C
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E
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I
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K
|
510 |
TM7 | |||||||||
N
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R
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P
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S
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L
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L
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V
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E
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K
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I
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520 |
N
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L
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F
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A
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M
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F
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G
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T
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G
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I
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530 |
H8 | |||||||||
A
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M
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S
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T
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W
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V
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W
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T
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K
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A
|
540 |
T
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L
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L
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I
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W
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R
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R
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T
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W
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C
|
550 |
|
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||||||||
C-term | |||||||||
R
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L
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T
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G
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Q
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S
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D
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D
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E
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P
|
560 |
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K
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R
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I
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K
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K
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S
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K
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M
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I
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A
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570 |
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K
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A
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F
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S
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K
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R
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H
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E
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L
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L
|
580 |
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|
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Q
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N
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P
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G
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Q
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E
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L
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S
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F
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S
|
590 |
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|
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M
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H
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T
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V
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S
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H
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D
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G
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P
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V
|
600 |
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|
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A
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G
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L
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A
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F
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D
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L
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N
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E
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P
|
610 |
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S
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A
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D
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V
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S
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S
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A
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W
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A
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Q
|
620 |
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H
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V
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T
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K
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M
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V
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A
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R
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R
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G
|
630 |
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A
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L
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P
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Q
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D
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I
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S
|
V
|
T
|
640 |
|
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P
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V
|
A
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|
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V
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P
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P
|
E
|
E
|
650 |
|
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|
|
|
|
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|
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Q
|
A
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N
|
L
|
W
|
L
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V
|
E
|
A
|
E
|
660 |
|
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I
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S
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P
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L
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Q
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K
|
R
|
L
|
G
|
670 |
|
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|
|
|
|
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R
|
K
|
K
|
K
|
R
|
R
|
K
|
R
|
K
|
K
|
680 |
|
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|
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E
|
V
|
C
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P
|
L
|
A
|
P
|
P
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P
|
E
|
690 |
|
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|
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|
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|
|
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L
|
H
|
P
|
P
|
A
|
P
|
A
|
P
|
S
|
T
|
700 |
|
|
|
|
|
|
|
|
|
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I
|
P
|
R
|
L
|
P
|
Q
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L
|
P
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R
|
Q
|
710 |
|
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|
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|
|
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K
|
C
|
L
|
V
|
A
|
A
|
G
|
A
|
W
|
G
|
720 |
|
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|
|
|
|
|
|
|
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A
|
G
|
D
|
S
|
C
|
R
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Q
|
G
|
A
|
W
|
730 |
|
|
|
|
|
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|
|
|
|
T
|
L
|
V
|
S
|
N
|
P
|
F
|
C
|
P
|
E
|
740 |
|
|
|
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|
|
|
|
|
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P
|
S
|
P
|
P
|
Q
|
D
|
P
|
F
|
L
|
P
|
750 |
|
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|
|
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|
|
|
|
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S
|
A
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P
|
A
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P
|
V
|
A
|
W
|
A
|
H
|
760 |
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G
|
R
|
R
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Q
|
G
|
L
|
G
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P
|
I
|
H
|
770 |
|
|
|
|
|
|
|
|
|
|
S
|
R
|
T
|
N
|
L
|
M
|
D
|
T
|
E
|
L
|
780 |
|
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|
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|
|
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M
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D
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A
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D
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S
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D
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F
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hemagglutin | ||||||||||
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FLAG | |||||||||
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His(10) | |||||||||
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TEV proteas | ||||||||||
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|||||||||
N-term | |||||||||
|
P
|
P
|
P
|
L
|
S
|
H
|
C
|
G
|
R
|
66 |
A
|
A
|
P
|
C
|
E
|
P
|
L
|
R
|
Y
|
N
|
76 |
V
|
C
|
L
|
G
|
S
|
V
|
L
|
P
|
Y
|
G
|
86 |
A
|
T
|
S
|
T
|
L
|
L
|
A
|
G
|
D
|
S
|
96 |
D
|
S
|
Q
|
E
|
E
|
A
|
H
|
G
|
K
|
L
|
106 |
V
|
L
|
W
|
S
|
G
|
L
|
R
|
N
|
A
|
P
|
116 |
R
|
C
|
W
|
A
|
V
|
I
|
Q
|
P
|
L
|
L
|
126 |
C
|
A
|
V
|
Y
|
M
|
P
|
K
|
C
|
E
|
N
|
136 |
D
|
R
|
V
|
E
|
L
|
P
|
S
|
R
|
T
|
L
|
146 |
C
|
Q
|
A
|
T
|
R
|
G
|
P
|
C
|
A
|
I
|
156 |
V
|
E
|
R
|
E
|
R
|
G
|
W
|
P
|
D
|
F
|
166 |
L
|
R
|
C
|
T
|
P
|
D
|
R
|
F
|
P
|
E
|
176 |
G
|
C
|
T
|
N
|
E
|
V
|
Q
|
N
|
I
|
K
|
186 |
F
|
N
|
S
|
S
|
G
|
Q
|
C
|
E
|
V
|
P
|
196 |
L
|
V
|
R
|
T
|
D
|
N
|
P
|
K
|
S
|
W
|
206 |
Y
|
E
|
D
|
V
|
E
|
G
|
C
|
G
|
I
|
Q
|
216 |
TM1 | |||||||||
C
|
Q
|
N
|
P
|
L
|
F
|
T
|
E
|
A
|
E
|
226 |
H
|
Q
|
D
|
M
|
H
|
S
|
Y
|
I
|
A
|
A
|
236 |
F
|
G
|
A
|
V
|
T
|
G
|
L
|
C
|
T
|
L
|
246 |
F
|
T
|
L
|
A
|
T
|
F
|
V
|
A
|
D
|
W
|
256 |
ICL1 | TM2 | ||||||||
R
|
N
|
S
|
N
|
R
|
Y
|
P
|
A
|
V
|
I
|
266 |
L
|
F
|
Y
|
V
|
N
|
A
|
C
|
F
|
F
|
V
|
276 |
G
|
S
|
I
|
G
|
W
|
L
|
A
|
Q
|
F
|
M
|
286 |
ECL1 | |||||||||
D
|
G
|
A
|
R
|
R
|
E
|
I
|
V
|
C
|
R
|
296 |
A
|
D
|
G
|
T
|
M
|
R
|
L
|
G
|
E
|
P
|
306 |
TM3 | |||||||||
T
|
S
|
N
|
E
|
T
|
L
|
S
|
C
|
V
|
I
|
316 |
I
|
F
|
V
|
I
|
V
|
Y
|
Y
|
A
|
L
|
M
|
326 |
A
|
G
|
V
|
V
|
W
|
F
|
V
|
V
|
L
|
T
|
336 |
Y
|
A
|
W
|
H
|
T
|
S
|
F
|
K
|
A
|
L
|
346 |
ICL2 | |||||||||
G
|
T
|
T
|
Y
|
Q
|
P
|
L
|
S
|
G
|
K
|
356 |
TM4 | |||||||||
T
|
S
|
Y
|
F
|
H
|
L
|
L
|
T
|
W
|
S
|
366 |
L
|
P
|
F
|
V
|
L
|
T
|
V
|
A
|
I
|
L
|
376 |
ECL2 | |||||||||
A
|
V
|
A
|
Q
|
V
|
D
|
G
|
D
|
S
|
V
|
386 |
S
|
G
|
I
|
C
|
F
|
V
|
G
|
Y
|
K
|
N
|
396 |
TM5 | |||||||||
Y
|
R
|
Y
|
R
|
A
|
G
|
F
|
V
|
L
|
A
|
406 |
P
|
I
|
G
|
L
|
V
|
L
|
I
|
V
|
G
|
G
|
416 |
Y
|
F
|
L
|
I
|
R
|
G
|
V
|
M
|
T
|
L
|
426 |
|
|||||||
F
|
S
|
I
|
K
|
S
|
N
|
H
|
|
|
|
|
|
|
|
|
|
|
|
Flavodoxin | |||||||||
|
|
|
|
|
|
|
|
|
|
TM6 | |||||||
K
|
I
|
N
|
E
|
T
|
M
|
L
|
R
|
451 |
L
|
G
|
I
|
F
|
G
|
F
|
L
|
A
|
F
|
G
|
461 |
F
|
V
|
L
|
I
|
T
|
F
|
S
|
C
|
H
|
F
|
471 |
Y
|
D
|
F
|
F
|
N
|
Q
|
A
|
E
|
W
|
E
|
481 |
R
|
S
|
F
|
R
|
D
|
Y
|
V
|
L
|
C
|
Q
|
491 |
ECL3 | |||||||||
A
|
N
|
V
|
T
|
I
|
G
|
L
|
P
|
T
|
K
|
501 |
Q
|
P
|
I
|
P
|
D
|
C
|
E
|
I
|
K
|
N
|
511 |
TM7 | |||||||||
R
|
P
|
S
|
L
|
L
|
V
|
E
|
K
|
I
|
N
|
521 |
L
|
F
|
A
|
M
|
F
|
G
|
T
|
G
|
I
|
A
|
531 |
H8 | |||||||||
M
|
S
|
T
|
W
|
V
|
W
|
T
|
K
|
A
|
T
|
541 |
L
|
L
|
I
|
W
|
R
|
R
|
T
|
W
|
C
|
R
|
551 |
|
|||||||
C-term | |||||||
L
|
T
|
G
|
Q
|
S
|
D
|
D
|
|
|
|
|
|
|
|
|
|
|
|
His(10) | |||||||||
|
|
|
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|
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|
|
SCHEMATIC
Perspective:
Wild-type
Construct
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DIAGRAMS
MUTATIONS
Position | Wild-type | Mutated |
---|
DELETIONS
MODIFICATIONS
AUXILIARY PROTEINS
Protein_type: signal
Name: hemagglutinin
Sequence: None
Deletions:
Positions: N-term_0 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: tag
Name: FLAG
Sequence: None
Deletions:
Positions: N-term_1 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: tag
Name: His(10)
Sequence: None
Deletions:
Positions: N-term_2 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: prot_cleavage
Name: TEV protease
Sequence: None
Deletions:
Positions: N-term_3 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: fusion
Name: Flavodoxin
Sequence: None
Deletions:
Positions: Within Receptor_4 434:443
Presence in Crystal:YES
Remarks: None
Protein_type: tag
Name: His(10)
Sequence: None
Deletions:
Positions: C-term_5 None:None
Presence in Crystal:YES
Remarks: None
EXPRESSION
Expression method | Host cell type | Host cell | Remarks |
---|---|---|---|
Transient_transfection | HEK293F | Mammalian | HEK293Fcells(Invitrogen)weregrowninsuspension + FreeStyleTM 293 Expression system |
SOLUBILIZATION
Solvent type | Item | Concentration |
---|---|---|
detergent | DDM | 0.2 %w/v |
additive | CHS | 1.2 %w/v |
Remarks:
PURIFICATION
Purification type | Description |
---|---|
Proteolysis by TEV protease | None |
Detergent DDM exchanged with LMNG | None |
Remarks:None
CRYSTALLIZATION
METHOD: in meso/LCP
TYPE: lipidic cubic phase (LCP) (monoolein (9.9 MAG))
PROTEIN CONCENTRATION: 40-50 mg/ml
Temperature: 20°C
ph : 5.0-5.0
Remarks: Solved at Synchrotron
Chemical lists
Type | Item | Concentration |
---|---|---|
Detergent | ||
LMNG | 0.01 %v/v | |
Lipid | ||
CHS | 0.002 % v/v | |
Additional | ||
Buffer | Sodium citrate tribasic dihydrate, pH 5.0 | 100.0 mM |
Precipitant | PEG 400 | 36.0 % v/v |
Ammonium nitrate | 50-200 mM |