adrb2_human
NAME
5X7D
ORGANISM
Human (Homo sapiens)
PROTEIN NAME
adrb2_human
SOURCE
SWISSPROT
SEQUENCE
Perspective:
Wild-type
Construct
N-term | |||||||||
M
|
G
|
Q
|
P
|
G
|
N
|
G
|
S
|
A
|
F
|
10 |
|
|||||||||
L
|
L
|
A
|
P
|
N
|
G
|
S
|
H
|
A
|
P
|
20 |
|
|||||||||
TM1 | |||||||||
D
|
H
|
D
|
V
|
T
|
Q
|
E
|
R
|
D
|
E
|
30 |
V
|
W
|
V
|
V
|
G
|
M
|
G
|
I
|
V
|
M
|
40 |
S
|
L
|
I
|
V
|
L
|
A
|
I
|
V
|
F
|
G
|
50 |
N
|
V
|
L
|
V
|
I
|
T
|
A
|
I
|
A
|
K
|
60 |
ICL1 | TM2 | ||||||||
F
|
E
|
R
|
L
|
Q
|
T
|
V
|
T
|
N
|
Y
|
70 |
F
|
I
|
T
|
S
|
L
|
A
|
C
|
A
|
D
|
L
|
80 |
V
|
M
|
G
|
L
|
A
|
V
|
V
|
P
|
F
|
G
|
90 |
A
|
A
|
H
|
I
|
L
|
M
|
K
|
M
|
W
|
T
|
100 |
ECL1 | TM3 | ||||||||
F
|
G
|
N
|
F
|
W
|
C
|
E
|
F
|
W
|
T
|
110 |
S
|
I
|
D
|
V
|
L
|
C
|
V
|
T
|
A
|
S
|
120 |
I
|
E
|
T
|
L
|
C
|
V
|
I
|
A
|
V
|
D
|
130 |
R
|
Y
|
F
|
A
|
I
|
T
|
S
|
P
|
F
|
K
|
140 |
ICL2 | TM4 | ||||||||
Y
|
Q
|
S
|
L
|
L
|
T
|
K
|
N
|
K
|
A
|
150 |
R
|
V
|
I
|
I
|
L
|
M
|
V
|
W
|
I
|
V
|
160 |
S
|
G
|
L
|
T
|
S
|
F
|
L
|
P
|
I
|
Q
|
170 |
ECL2 | |||||||||
M
|
H
|
W
|
Y
|
R
|
A
|
T
|
H
|
Q
|
E
|
180 |
|
|||||||||
A
|
I
|
N
|
C
|
Y
|
A
|
N
|
E
|
T
|
C
|
190 |
TM5 | |||||||||
C
|
D
|
F
|
F
|
T
|
N
|
Q
|
A
|
Y
|
A
|
200 |
I
|
A
|
S
|
S
|
I
|
V
|
S
|
F
|
Y
|
V
|
210 |
P
|
L
|
V
|
I
|
M
|
V
|
F
|
V
|
Y
|
S
|
220 |
R
|
V
|
F
|
Q
|
E
|
A
|
K
|
R
|
Q
|
L
|
230 |
|
|
|
|
|
|
|
|
|
|
Q
|
K
|
I
|
D
|
K
|
S
|
E
|
G
|
R
|
F
|
240 |
|
|
|
|
|
|
|
|
|
|
ICL3 | |||||||||
H
|
V
|
Q
|
N
|
L
|
S
|
Q
|
V
|
E
|
Q
|
250 |
|
|
|
|
|
|
|
|
|
|
D
|
G
|
R
|
T
|
G
|
H
|
G
|
L
|
R
|
R
|
260 |
|
|
|
|||||||
TM6 | |||||||||
S
|
S
|
K
|
F
|
C
|
L
|
K
|
E
|
H
|
K
|
270 |
A
|
L
|
K
|
T
|
L
|
G
|
I
|
I
|
M
|
G
|
280 |
T
|
F
|
T
|
L
|
C
|
W
|
L
|
P
|
F
|
F
|
290 |
I
|
V
|
N
|
I
|
V
|
H
|
V
|
I
|
Q
|
D
|
300 |
ECL3 | TM7 | ||||||||
N
|
L
|
I
|
R
|
K
|
E
|
V
|
Y
|
I
|
L
|
310 |
L
|
N
|
W
|
I
|
G
|
Y
|
V
|
N
|
S
|
G
|
320 |
H8 | |||||||||
F
|
N
|
P
|
L
|
I
|
Y
|
C
|
R
|
S
|
P
|
330 |
D
|
F
|
R
|
I
|
A
|
F
|
Q
|
E
|
L
|
L
|
340 |
C-term | |||||||||
C
|
L
|
R
|
R
|
S
|
S
|
L
|
K
|
A
|
Y
|
350 |
G
|
N
|
G
|
Y
|
S
|
S
|
N
|
G
|
N
|
T
|
360 |
|
|
|
|
|
|||||
G
|
E
|
Q
|
S
|
G
|
Y
|
H
|
V
|
E
|
Q
|
370 |
|
|
|
|
|
|
|
|
|
|
E
|
K
|
E
|
N
|
K
|
L
|
L
|
C
|
E
|
D
|
380 |
|
|
|
|
|
|
|
|
|
|
L
|
P
|
G
|
T
|
E
|
D
|
F
|
V
|
G
|
H
|
390 |
|
|
|
|
|
|
|
|
|
|
Q
|
G
|
T
|
V
|
P
|
S
|
D
|
N
|
I
|
D
|
400 |
|
|
|
|
|
|
|
|
|
|
S
|
Q
|
G
|
R
|
N
|
C
|
S
|
T
|
N
|
D
|
410 |
|
|
|
S
|
L
|
L
|
|
|
|
|
|
|
|
|
|
|
FLAG | |||||||||
|
|
|
|
|
|
|
|
|
|
N-term | |||||||||
M
|
G
|
Q
|
P
|
G
|
N
|
G
|
S
|
A
|
F
|
10 |
|
|||||||||
L
|
L
|
A
|
P
|
N
|
R
|
S
|
H
|
A
|
P
|
20 |
|
|||||||||
TM1 | |||||||||
D
|
H
|
D
|
V
|
T
|
Q
|
Q
|
R
|
D
|
E
|
30 |
V
|
W
|
V
|
V
|
G
|
M
|
G
|
I
|
V
|
M
|
40 |
S
|
L
|
I
|
V
|
L
|
A
|
I
|
V
|
F
|
G
|
50 |
N
|
V
|
L
|
V
|
I
|
T
|
A
|
I
|
A
|
K
|
60 |
ICL1 | TM2 | ||||||||
F
|
E
|
R
|
L
|
Q
|
T
|
V
|
T
|
N
|
Y
|
70 |
F
|
I
|
T
|
S
|
L
|
A
|
C
|
A
|
D
|
L
|
80 |
V
|
M
|
G
|
L
|
A
|
V
|
V
|
P
|
F
|
G
|
90 |
A
|
A
|
H
|
I
|
L
|
M
|
K
|
M
|
W
|
T
|
100 |
ECL1 | TM3 | ||||||||
F
|
G
|
N
|
F
|
W
|
C
|
E
|
F
|
W
|
T
|
110 |
S
|
I
|
D
|
V
|
L
|
C
|
V
|
T
|
A
|
S
|
120 |
I
|
E
|
T
|
L
|
C
|
V
|
I
|
A
|
V
|
D
|
130 |
R
|
Y
|
F
|
A
|
I
|
T
|
S
|
P
|
F
|
K
|
140 |
ICL2 | TM4 | ||||||||
Y
|
Q
|
S
|
L
|
L
|
T
|
K
|
N
|
K
|
A
|
150 |
R
|
V
|
I
|
I
|
L
|
M
|
V
|
W
|
I
|
V
|
160 |
S
|
G
|
L
|
T
|
S
|
F
|
L
|
P
|
I
|
Q
|
170 |
ECL2 | |||||||||
M
|
H
|
W
|
Y
|
R
|
A
|
T
|
H
|
Q
|
E
|
180 |
|
|||||||||
A
|
I
|
N
|
C
|
Y
|
A
|
E
|
E
|
T
|
C
|
190 |
TM5 | |||||||||
C
|
D
|
F
|
F
|
T
|
N
|
Q
|
A
|
Y
|
A
|
200 |
I
|
A
|
S
|
S
|
I
|
V
|
S
|
F
|
Y
|
V
|
210 |
P
|
L
|
V
|
I
|
M
|
V
|
F
|
V
|
Y
|
S
|
220 |
|
|||||||||||||||||||||||||||||||||||||||
TM6 | |||||||||||||||||||||||||||||||||||||||
R
|
V
|
F
|
Q
|
E
|
A
|
K
|
R
|
Q
|
L
|
|
F
|
C
|
L
|
K
|
E
|
H
|
K
|
A
|
L
|
||||||||||||||||||||
230 | 272 |
K
|
T
|
L
|
G
|
I
|
I
|
M
|
G
|
T
|
F
|
282 |
T
|
L
|
C
|
W
|
L
|
P
|
F
|
F
|
I
|
V
|
292 |
N
|
I
|
V
|
H
|
V
|
I
|
Q
|
D
|
N
|
L
|
302 |
ECL3 | TM7 | ||||||||
I
|
R
|
K
|
E
|
V
|
Y
|
I
|
L
|
L
|
N
|
312 |
W
|
I
|
G
|
Y
|
V
|
N
|
S
|
G
|
F
|
N
|
322 |
H8 | |||||||||
P
|
L
|
I
|
Y
|
C
|
R
|
S
|
P
|
D
|
F
|
332 |
R
|
I
|
A
|
F
|
Q
|
E
|
L
|
L
|
C
|
L
|
342 |
C-term | |||||||||
R
|
R
|
S
|
S
|
L
|
K
|
A
|
Y
|
G
|
N
|
352 |
G
|
Y
|
S
|
S
|
N
|
G
|
N
|
T
|
G
|
E
|
362 |
|
|||
Q
|
S
|
G
|
|
SCHEMATIC
Perspective:
Wild-type
Construct
|
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DIAGRAMS
MUTATIONS
Position | Wild-type | Mutated |
---|---|---|
16 | | G | R |
27 | | E | Q |
187 | | N | E |
DELETIONS
MODIFICATIONS
AUXILIARY PROTEINS
Protein_type: tag
Name: FLAG
Sequence: None
Deletions:
Positions: N-term_0 None:None
Presence in Crystal:YES
Remarks: None
Protein_type: fusion
Name: T4 Lysozyme (T4L)
Sequence: None
Deletions:
Positions: Within Receptor_1 233:260
Presence in Crystal:YES
Remarks: None
EXPRESSION
Expression method | Host cell type | Host cell | Remarks |
---|---|---|---|
Baculovirus | Sf9 | Insect |
SOLUBILIZATION
Solvent type | Item | Concentration |
---|---|---|
detergent | DDM | 1.0 %w/v |
Remarks:
PURIFICATION
Purification type | Description |
---|---|
Alkylation by Iodoacetamide | None |
Deglycosylation by PNGase F | None |
detergent was exchanged from 0.1% DDM to 0.01% MNG | None |
Remarks:None
CRYSTALLIZATION
METHOD: in meso/LCP
TYPE: lipidic cubic phase (LCP) (monoolein (9.9 MAG))
PROTEIN CONCENTRATION: 50 mg/ml
Temperature: 20°C
ph : 7.5-7.5
Remarks: PEG 400 and sulfate ion were essential for crystallization, while addition of cholesterol and 1,4-butanediol dramatically improved crystals size and shape enabling high-resolution diffraction. Additions of phospholipids(dioleoylphosphatidylcholine, dioleoylphosphatidylethanolamine, asolectin) alone and in combinations with cholesterol to the main host LCP lipid monoolein were tried, however, none of them improved crystal quality. 52-50% Monoolein96-well glass sandwich plates (S1, S2) were filled with 25 or 50 nLprotein-laden LCP drops overlaid by 0.8 μL of precipitant solution in each well
Chemical lists
Type | Item | Concentration |
---|---|---|
Buffer | ||
Tris-HCl pH 7.5 | 0.1 M | |
Additional | ||
PEG400 | 30–37.5 %v/v | |
NH4F | 400.0 mM | |
1,4-butanediol | 6.0 %v/v | |
DMSO. | 1.0 % | |
Detergent | ||
LMNG | 0.003 % | |
Lipid | ||
CHS | 0.0003 % |