oprm_mouse
NAME
6DDE
ORGANISM
Mouse (Mus musculus)
PROTEIN NAME
oprm_mouse
SOURCE
SWISSPROT
SEQUENCE
Perspective:
Wild-type
Construct
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N-term | |||||||||
M
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D
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S
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S
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A
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G
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P
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G
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N
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I
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10 |
S
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D
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C
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S
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D
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P
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L
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A
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P
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A
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20 |
S
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C
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S
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P
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A
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P
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G
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S
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W
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L
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30 |
N
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L
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S
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H
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V
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D
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G
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N
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Q
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S
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40 |
D
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P
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C
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G
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P
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N
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R
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T
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G
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L
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50 |
G
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G
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S
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H
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S
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L
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C
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P
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Q
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T
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60 |
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|||||||||
TM1 | |||||||||
G
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S
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P
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S
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M
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V
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T
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A
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I
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T
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70 |
I
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M
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A
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L
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Y
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S
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I
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V
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C
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V
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80 |
V
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G
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L
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F
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G
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N
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F
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L
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V
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M
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90 |
ICL1 | |||||||||
Y
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V
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I
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V
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R
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Y
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T
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K
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M
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K
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100 |
TM2 | |||||||||
T
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A
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T
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N
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I
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Y
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I
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F
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N
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L
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110 |
A
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L
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A
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D
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A
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L
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A
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T
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S
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T
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120 |
L
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P
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F
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Q
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S
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V
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N
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Y
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L
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M
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130 |
ECL1 | TM3 | ||||||||
G
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T
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W
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P
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F
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G
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N
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I
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L
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C
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140 |
K
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I
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V
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I
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S
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I
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D
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Y
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Y
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N
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150 |
M
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F
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T
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S
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I
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F
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T
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L
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C
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T
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160 |
M
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S
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V
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D
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R
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Y
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I
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A
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V
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C
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170 |
ICL2 | |||||||||
H
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P
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V
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K
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A
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L
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D
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F
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R
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T
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180 |
TM4 | |||||||||
P
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R
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N
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A
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K
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I
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V
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N
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V
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C
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190 |
N
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W
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I
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L
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S
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S
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A
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I
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G
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L
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200 |
ECL2 | |||||||||
P
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V
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M
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F
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M
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A
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T
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T
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K
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Y
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210 |
R
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Q
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G
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S
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I
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D
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C
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T
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L
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T
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220 |
TM5 | |||||||||
F
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S
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H
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P
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T
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W
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Y
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W
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E
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N
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230 |
L
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L
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K
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I
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C
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V
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F
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I
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F
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A
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240 |
F
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I
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M
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P
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V
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L
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I
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I
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T
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V
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250 |
C
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Y
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G
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L
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M
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I
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L
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R
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L
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K
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260 |
ICL3 | TM6 | ||||||||
S
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V
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R
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M
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L
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S
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G
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S
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K
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E
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270 |
K
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D
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R
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N
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L
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R
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R
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I
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T
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R
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280 |
M
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V
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L
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V
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V
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V
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A
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V
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F
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I
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290 |
V
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C
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W
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T
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P
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I
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H
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I
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Y
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V
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300 |
ECL3 | |||||||||
I
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I
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K
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A
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L
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I
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T
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I
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P
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E
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310 |
TM7 | |||||||||
T
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T
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F
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Q
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T
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V
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S
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W
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H
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F
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320 |
C
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I
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A
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L
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G
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Y
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T
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N
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S
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C
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330 |
L
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N
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P
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V
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L
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Y
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A
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F
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L
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D
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340 |
H8 | |||||||||
E
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N
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F
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K
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R
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C
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F
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R
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E
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F
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350 |
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C-term | |||||||||
C
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I
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P
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T
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S
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S
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T
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I
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E
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Q
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360 |
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|
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Q
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N
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S
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A
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R
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I
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R
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Q
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N
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T
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370 |
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|
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R
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E
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H
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P
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S
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T
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A
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N
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T
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V
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380 |
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|
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D
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R
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T
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N
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H
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Q
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L
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E
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N
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L
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390 |
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E
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A
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E
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T
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A
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P
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L
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P
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FLAG | |||||||||
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N-term | |||||||||
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N
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I
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S
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D
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C
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S
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D
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P
|
L
|
17 |
A
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P
|
A
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S
|
C
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S
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P
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A
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P
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G
|
27 |
S
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W
|
L
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N
|
L
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S
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H
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V
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D
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G
|
37 |
N
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Q
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S
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D
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P
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C
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G
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P
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N
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R
|
47 |
T
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G
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L
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G
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G
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S
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H
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S
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L
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C
|
57 |
P
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Q
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T
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G
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S
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P
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63 |
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TEV proteas | ||||||||||
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TM1 | ||||||||
S
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M
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V
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T
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A
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I
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T
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I
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M
|
72 |
A
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L
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Y
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S
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I
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V
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C
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V
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V
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G
|
82 |
L
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F
|
G
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N
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F
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L
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V
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M
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Y
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V
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92 |
ICL1 | |||||||||
I
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V
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R
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Y
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T
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K
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M
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K
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T
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A
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102 |
TM2 | |||||||||
T
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N
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I
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Y
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I
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F
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N
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L
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A
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L
|
112 |
A
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D
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A
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L
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A
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T
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S
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T
|
L
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P
|
122 |
F
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Q
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S
|
V
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N
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Y
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L
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M
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G
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T
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132 |
ECL1 | TM3 | ||||||||
W
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P
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F
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G
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N
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I
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L
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C
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K
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I
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142 |
V
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I
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S
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I
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D
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Y
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Y
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N
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M
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F
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152 |
T
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S
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I
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F
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T
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L
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C
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T
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M
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S
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162 |
V
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D
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R
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Y
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I
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A
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V
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C
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H
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P
|
172 |
ICL2 | TM4 | ||||||||
V
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K
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A
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L
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D
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F
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R
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T
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P
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R
|
182 |
N
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A
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K
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I
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V
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N
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V
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C
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N
|
W
|
192 |
I
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L
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S
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S
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A
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I
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G
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L
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P
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V
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202 |
ECL2 | |||||||||
M
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F
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M
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A
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T
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T
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K
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Y
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R
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Q
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212 |
G
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S
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I
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D
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C
|
T
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L
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T
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F
|
S
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222 |
TM5 | |||||||||
H
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P
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T
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W
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Y
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W
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E
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N
|
L
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L
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232 |
K
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I
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C
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V
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F
|
I
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F
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A
|
F
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I
|
242 |
M
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P
|
V
|
L
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I
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I
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T
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V
|
C
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Y
|
252 |
G
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L
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M
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I
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L
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R
|
L
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K
|
S
|
V
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262 |
ICL3 | TM6 | ||||||||
R
|
M
|
L
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S
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G
|
S
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K
|
E
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K
|
D
|
272 |
R
|
N
|
L
|
R
|
R
|
I
|
T
|
R
|
M
|
V
|
282 |
L
|
V
|
V
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V
|
A
|
V
|
F
|
I
|
V
|
C
|
292 |
W
|
T
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P
|
I
|
H
|
I
|
Y
|
V
|
I
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I
|
302 |
ECL3 | |||||||||
K
|
A
|
L
|
I
|
T
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I
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P
|
E
|
T
|
T
|
312 |
TM7 | |||||||||
F
|
Q
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T
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V
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S
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W
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H
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F
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C
|
I
|
322 |
A
|
L
|
G
|
Y
|
T
|
N
|
S
|
C
|
L
|
N
|
332 |
H8 | |||||||||
P
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V
|
L
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Y
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A
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F
|
L
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D
|
E
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N
|
342 |
F
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K
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R
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C
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F
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R
|
E
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F
|
C
|
I
|
352 |
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C-term | ||||||
P
|
T
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S
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S
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T
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I
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PreScission | ||||||||||
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PreScission | ||||||||||
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His(6) | |||||||||
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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: tag
Name: FLAG
Sequence: None
Deletions:
Positions: N-term_0 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: prot_cleavage
Name: TEV protease
Sequence: None
Deletions:
Positions: Within Receptor_0 64:64
Presence in Crystal:YES
Remarks: None
Protein_type: prot_cleavage
Name: PreScission site
Sequence: None
Deletions:
Positions: C-term_2 359:359
Presence in Crystal:NO
Remarks: None
Protein_type: tag
Name: His(6)
Sequence: None
Deletions:
Positions: C-term_3 None:None
Presence in Crystal:NO
Remarks: None
EXPRESSION
Expression method | Host cell type | Host cell | Remarks |
---|---|---|---|
Baculovirus | Sf9 | Insect | BestBac |
SOLUBILIZATION
Solvent type | Item | Concentration |
---|---|---|
detergent | LMNG | 0.1 %w/v |
additive | CHS | 0.001 %w/v |
Remarks:
PURIFICATION
Purification type | Description |
---|---|
Proteolysis by TEV protease | None |
Proteolysis by HRV 3C protease | None |
Remarks:None
CRYSTALLIZATION
METHOD: Cryo-EM
TYPE: Gatan K2 Summit direct electron camera , 300kV ()
PROTEIN CONCENTRATION: 7.0 mg/ml
Temperature: 4°C
ph : 7.5-7.5
Remarks: nucleotide free Gi, with Fab scFv16
Chemical lists
Type | Item | Concentration |
---|---|---|
Detergent | ||
LMNG | 0.00075 %v/v | |
Lipid | ||
CHS | 0.0001 %v/v | |
Additional | ||
Buffer | Hepes pH 7.5 | 20.0 mM |
NaCl | 100.0 mM | |
glyco-diosgenin (GDN) | 0.00025 %v/v |