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
|
A
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G
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P
|
G
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N
|
I
|
10 |
S
|
D
|
C
|
S
|
D
|
P
|
L
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A
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P
|
A
|
20 |
S
|
C
|
S
|
P
|
A
|
P
|
G
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S
|
W
|
L
|
30 |
N
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L
|
S
|
H
|
V
|
D
|
G
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N
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Q
|
S
|
40 |
D
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P
|
C
|
G
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P
|
N
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R
|
T
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G
|
L
|
50 |
G
|
G
|
S
|
H
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S
|
L
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C
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P
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Q
|
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
|
T
|
70 |
I
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M
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A
|
L
|
Y
|
S
|
I
|
V
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C
|
V
|
80 |
V
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G
|
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
|
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
|
L
|
110 |
A
|
L
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A
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D
|
A
|
L
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A
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T
|
S
|
T
|
120 |
L
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P
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F
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Q
|
S
|
V
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N
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Y
|
L
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M
|
130 |
ECL1 | TM3 | ||||||||
G
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T
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W
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P
|
F
|
G
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N
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I
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L
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C
|
140 |
K
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I
|
V
|
I
|
S
|
I
|
D
|
Y
|
Y
|
N
|
150 |
M
|
F
|
T
|
S
|
I
|
F
|
T
|
L
|
C
|
T
|
160 |
M
|
S
|
V
|
D
|
R
|
Y
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I
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A
|
V
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C
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170 |
ICL2 | |||||||||
H
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P
|
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
|
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
|
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
|
I
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G
|
L
|
200 |
ECL2 | |||||||||
P
|
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
|
210 |
R
|
Q
|
G
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S
|
I
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D
|
C
|
T
|
L
|
T
|
220 |
TM5 | |||||||||
F
|
S
|
H
|
P
|
T
|
W
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Y
|
W
|
E
|
N
|
230 |
L
|
L
|
K
|
I
|
C
|
V
|
F
|
I
|
F
|
A
|
240 |
F
|
I
|
M
|
P
|
V
|
L
|
I
|
I
|
T
|
V
|
250 |
C
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Y
|
G
|
L
|
M
|
I
|
L
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R
|
L
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K
|
260 |
ICL3 | TM6 | ||||||||
S
|
V
|
R
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M
|
L
|
S
|
G
|
S
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K
|
E
|
270 |
K
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D
|
R
|
N
|
L
|
R
|
R
|
I
|
T
|
R
|
280 |
M
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V
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L
|
V
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V
|
V
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A
|
V
|
F
|
I
|
290 |
V
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C
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W
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T
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P
|
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
|
E
|
310 |
TM7 | |||||||||
T
|
T
|
F
|
Q
|
T
|
V
|
S
|
W
|
H
|
F
|
320 |
C
|
I
|
A
|
L
|
G
|
Y
|
T
|
N
|
S
|
C
|
330 |
L
|
N
|
P
|
V
|
L
|
Y
|
A
|
F
|
L
|
D
|
340 |
H8 | |||||||||
E
|
N
|
F
|
K
|
R
|
C
|
F
|
R
|
E
|
F
|
350 |
|
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||||||||
C-term | |||||||||
C
|
I
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P
|
T
|
S
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S
|
T
|
I
|
E
|
Q
|
360 |
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|
|
|
|
|
|
|
|
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Q
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N
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S
|
A
|
R
|
I
|
R
|
Q
|
N
|
T
|
370 |
|
|
|
|
|
|
|
|
|
|
R
|
E
|
H
|
P
|
S
|
T
|
A
|
N
|
T
|
V
|
380 |
|
|
|
|
|
|
|
|
|
|
D
|
R
|
T
|
N
|
H
|
Q
|
L
|
E
|
N
|
L
|
390 |
|
|
|
|
|
|
|
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E
|
A
|
E
|
T
|
A
|
P
|
L
|
P
|
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 |