aa2ar_human
NAME
4UG2
ORGANISM
Human (Homo sapiens)
PROTEIN NAME
aa2ar_human
SOURCE
SWISSPROT
SEQUENCE
Perspective:
Wild-type
Construct
TM1 | |||||||||
M
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P
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I
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M
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G
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S
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S
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V
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Y
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I
|
10 |
T
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V
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E
|
L
|
A
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I
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A
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V
|
L
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A
|
20 |
I
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L
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G
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N
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V
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L
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V
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C
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W
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A
|
30 |
ICL1 | |||||||||
V
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W
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L
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N
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S
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N
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L
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Q
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N
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V
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40 |
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|||||||||
TM2 | |||||||||
T
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N
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Y
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F
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V
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V
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S
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L
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A
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A
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50 |
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|||||||||
A
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D
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I
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A
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V
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G
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V
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L
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A
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I
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60 |
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|||||||||
P
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F
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A
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I
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T
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I
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S
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T
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G
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F
|
70 |
ECL1 | TM3 | ||||||||
C
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A
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A
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C
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H
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G
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C
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L
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F
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I
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80 |
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|||||||||
A
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C
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F
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V
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L
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V
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L
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T
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Q
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S
|
90 |
S
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I
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F
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S
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L
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L
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A
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I
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A
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I
|
100 |
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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I
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R
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I
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P
|
L
|
110 |
ICL2 | TM4 | ||||||||
R
|
Y
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N
|
G
|
L
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V
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T
|
G
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T
|
R
|
120 |
A
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K
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G
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I
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I
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A
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I
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C
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W
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V
|
130 |
L
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S
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F
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A
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I
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G
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L
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T
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P
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M
|
140 |
ECL2 | |||||||||
L
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G
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W
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N
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N
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C
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G
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Q
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P
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K
|
150 |
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|||||||||
E
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G
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K
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N
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H
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S
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Q
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G
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C
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G
|
160 |
E
|
G
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Q
|
V
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A
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C
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L
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F
|
E
|
D
|
170 |
TM5 | |||||||||
V
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V
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P
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M
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N
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Y
|
M
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V
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Y
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F
|
180 |
N
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F
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F
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A
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C
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V
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L
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V
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P
|
L
|
190 |
L
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L
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M
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L
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G
|
V
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Y
|
L
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R
|
I
|
200 |
F
|
L
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A
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A
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R
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R
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Q
|
L
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K
|
Q
|
210 |
ICL3 | |||||||||
M
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E
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S
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Q
|
P
|
L
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P
|
G
|
E
|
R
|
220 |
TM6 | |||||||||
A
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R
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S
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T
|
L
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Q
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K
|
E
|
V
|
H
|
230 |
A
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A
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K
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S
|
L
|
A
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I
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I
|
V
|
G
|
240 |
L
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F
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A
|
L
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C
|
W
|
L
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P
|
L
|
H
|
250 |
I
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I
|
N
|
C
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F
|
T
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F
|
F
|
C
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P
|
260 |
ECL3 | TM7 | ||||||||
D
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C
|
S
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H
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A
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P
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L
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W
|
L
|
M
|
270 |
Y
|
L
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A
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I
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V
|
L
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S
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H
|
T
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N
|
280 |
S
|
V
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V
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N
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P
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F
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I
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Y
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A
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Y
|
290 |
H8 | |||||||||
R
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I
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R
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E
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F
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R
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Q
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T
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F
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R
|
300 |
K
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I
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I
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R
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S
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H
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V
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L
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R
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Q
|
310 |
|
|
|
|||||||
C-term | |||||||||
Q
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E
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P
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F
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K
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A
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A
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G
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T
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S
|
320 |
|
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|
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A
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R
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V
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L
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A
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A
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H
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G
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S
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D
|
330 |
|
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|
|
|
|
|
|
|
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G
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E
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Q
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V
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S
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L
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R
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L
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N
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G
|
340 |
|
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|
|
|
|
|
|
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H
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P
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P
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G
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V
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W
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A
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N
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G
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S
|
350 |
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A
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P
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H
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P
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E
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R
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R
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P
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N
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G
|
360 |
|
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|
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|
|
|
|
|
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Y
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A
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L
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G
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L
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V
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S
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G
|
G
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S
|
370 |
|
|
|
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|
|
|
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|
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A
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Q
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E
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S
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Q
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G
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N
|
T
|
G
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L
|
380 |
|
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|
|
|
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|
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|
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P
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D
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V
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E
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L
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L
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S
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H
|
E
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L
|
390 |
|
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|
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|
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K
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G
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V
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C
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P
|
E
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P
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P
|
G
|
L
|
400 |
|
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|
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|
|
|
|
|
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D
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D
|
P
|
L
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A
|
Q
|
D
|
G
|
A
|
G
|
410 |
|
|
V
|
S
|
TM1 | |||||||||
M
|
P
|
I
|
M
|
G
|
S
|
S
|
V
|
Y
|
I
|
10 |
T
|
V
|
E
|
L
|
A
|
I
|
A
|
V
|
L
|
A
|
20 |
I
|
L
|
G
|
N
|
V
|
L
|
V
|
C
|
W
|
A
|
30 |
ICL1 | |||||||||
V
|
W
|
L
|
N
|
S
|
N
|
L
|
Q
|
N
|
V
|
40 |
|
|||||||||
TM2 | |||||||||
T
|
N
|
Y
|
F
|
V
|
V
|
S
|
A
|
A
|
A
|
50 |
|
|||||||||
A
|
D
|
I
|
L
|
V
|
G
|
V
|
L
|
A
|
I
|
60 |
|
|||||||||
P
|
F
|
A
|
I
|
A
|
I
|
S
|
T
|
G
|
F
|
70 |
ECL1 | TM3 | ||||||||
C
|
A
|
A
|
C
|
H
|
G
|
C
|
L
|
F
|
I
|
80 |
|
|||||||||
A
|
C
|
F
|
V
|
L
|
V
|
L
|
T
|
A
|
S
|
90 |
S
|
I
|
F
|
S
|
L
|
L
|
A
|
I
|
A
|
I
|
100 |
D
|
R
|
Y
|
I
|
A
|
I
|
R
|
I
|
P
|
L
|
110 |
ICL2 | TM4 | ||||||||
R
|
Y
|
N
|
G
|
L
|
V
|
T
|
G
|
T
|
R
|
120 |
A
|
K
|
G
|
I
|
I
|
A
|
I
|
C
|
W
|
V
|
130 |
L
|
S
|
F
|
A
|
I
|
G
|
L
|
T
|
P
|
M
|
140 |
ECL2 | |||||||||
L
|
G
|
W
|
N
|
N
|
C
|
G
|
Q
|
P
|
K
|
150 |
|
|||||||||
E
|
G
|
K
|
A
|
H
|
S
|
Q
|
G
|
C
|
G
|
160 |
E
|
G
|
Q
|
V
|
A
|
C
|
L
|
F
|
E
|
D
|
170 |
TM5 | |||||||||
V
|
V
|
P
|
M
|
N
|
Y
|
M
|
V
|
Y
|
F
|
180 |
N
|
F
|
F
|
A
|
C
|
V
|
L
|
V
|
P
|
L
|
190 |
L
|
L
|
M
|
L
|
G
|
V
|
Y
|
L
|
R
|
I
|
200 |
F
|
L
|
A
|
A
|
R
|
R
|
Q
|
L
|
K
|
Q
|
210 |
ICL3 | |||||||||
M
|
E
|
S
|
Q
|
P
|
L
|
P
|
G
|
E
|
R
|
220 |
TM6 | |||||||||
A
|
R
|
S
|
T
|
L
|
Q
|
K
|
E
|
V
|
H
|
230 |
A
|
A
|
K
|
S
|
L
|
A
|
I
|
I
|
V
|
G
|
240 |
L
|
F
|
A
|
L
|
C
|
W
|
L
|
P
|
L
|
H
|
250 |
I
|
I
|
N
|
C
|
F
|
T
|
F
|
F
|
C
|
P
|
260 |
ECL3 | TM7 | ||||||||
D
|
C
|
S
|
H
|
A
|
P
|
L
|
W
|
L
|
M
|
270 |
Y
|
L
|
A
|
I
|
V
|
L
|
S
|
H
|
T
|
N
|
280 |
S
|
V
|
V
|
N
|
P
|
F
|
I
|
Y
|
A
|
Y
|
290 |
H8 | |||||||||
R
|
I
|
R
|
E
|
F
|
R
|
Q
|
T
|
F
|
R
|
300 |
K
|
I
|
I
|
R
|
S
|
H
|
V
|
L
|
R
|
Q
|
310 |
|
|||||||||
C-term | |||||||||
Q
|
E
|
P
|
F
|
K
|
A
|
A
|
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||
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AAA | |||||||||
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TEV proteas | ||||||||||
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His(10) | |||||||||
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SCHEMATIC
Perspective:
Wild-type
Construct
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DIAGRAMS
MUTATIONS
Position | Wild-type | Mutated |
---|---|---|
48 | | L | A |
54 | | A | L |
65 | | T | A |
89 | | Q | A |
154 | | N | A |
DELETIONS
MODIFICATIONS
AUXILIARY PROTEINS
Protein_type: prot_cleavage
Name: TEV protease
Sequence: None
Deletions:
Positions: C-term_1 None:None
Presence in Crystal:YES
Remarks: None
Protein_type: tag
Name: His(10)
Sequence: None
Deletions:
Positions: C-term_2 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: linker
Name: AAA
Sequence: None
Deletions:
Positions: C-term_0 None:None
Presence in Crystal:YES
Remarks: None
EXPRESSION
Expression method | Host cell type | Host cell | Remarks |
---|---|---|---|
Baculovirus | Hi5 | Insect | pBacPAK8 co-transfection linearized baculovirus |
SOLUBILIZATION
Solvent type | Item | Concentration |
---|---|---|
detergent | DM | 0.15 %w/v |
Remarks:
PURIFICATION
Purification type | Description |
---|---|
Proteolysis by TEV protease | None |
detergent exchange into 0.35% octylthioglucoside (OTG) | None |
Remarks:None
CRYSTALLIZATION
METHOD: in meso/LCP
TYPE: lipidic cubic phase (LCP) (monoolein (9.9 MAG))
PROTEIN CONCENTRATION: 3.5-4 mg/ml
Temperature: 22°C
ph : 7.0-7.0
Remarks: 2:3 (v/v) receptor-to-monoolein ratio, 0.15 % DM in monoolein + 10% cholesterol Purified receptor fractions were concen-trated to 15-25 mg/ml in a final volume of 50-60 μl
Chemical lists
Type | Item | Concentration |
---|---|---|
Additional | ||
unknown | ADA pH 7.0 | 0.1 M |
unknown | PEG 600 | 21.6 %v/v |
Detergent | ||
detergent | DM | 0.15 %w/v |