GPCRdb

agtr1_human

NAME

4ZUD

ORGANISM

Human (Homo sapiens)

PROTEIN NAME

agtr1_human

SOURCE

SWISSPROT

SEQUENCE

Perspective: Wild-type Construct


N-term
M	I	L	N	S	S	T	E	D	G
10



I	K	R	I	Q	D	D	C	P	K
20


			TM1
A	G	R	H	N	Y	I	F	V	M
30



I	P	T	L	Y	S	I	I	F	V
40



V	G	I	F	G	N	S	L	V	V
50


						ICL1
I	V	I	Y	F	Y	M	K	L	K
60


TM2
T	V	A	S	V	F	L	L	N	L
70



A	L	A	D	L	C	F	L	L	T
80



L	P	L	W	A	V	Y	T	A	M
90


	ECL1					TM3
E	Y	R	W	P	F	G	N	Y	L
100



C	K	I	A	S	A	S	V	S	F
110



N	L	Y	A	S	V	F	L	L	T
120



C	L	S	I	D	R	Y	L	A	I
130


		ICL2
V	H	P	M	K	S	R	L	R	R
140


TM4
T	M	L	V	A	K	V	T	C	I
150



I	I	W	L	L	A	G	L	A	S
160



L	P	A	I	I	H	R	N	V	F
170


ECL2
F	I	E	N	T	N	I	T	V	C
180



A	F	H	Y	E	S	Q	N	S	T
190


TM5
L	P	I	G	L	G	L	T	K	N
200



I	L	G	F	L	F	P	F	L	I
210



I	L	T	S	Y	T	L	I	W	K
220


					ICL3
A	L	K	K	A	Y	E	I	Q	K
230


		TM6
N	K	P	R	N	D	D	I	F	K
240



I	I	M	A	I	V	L	F	F	F
250



F	S	W	I	P	H	Q	I	F	T
260



F	L	D	V	L	I	Q	L	G	I
270


ECL3				TM7
I	R	D	C	R	I	A	D	I	V
280



D	T	A	M	P	I	T	I	C	I
290



A	Y	F	N	N	C	L	N	P	L
300


					H8
F	Y	G	F	L	G	K	K	F	K
310



R	Y	F	L	Q	L	L	K	Y	I
320


C-term
P	P	K	A	K	S	H	S	N	L
330



S	T	K	M	S	T	L	S	Y	R
340



P	S	D	N	V	S	S	S	T	K
350



K	P	A	P	C	F	E	V	E


hemagglutin


FLAG


BRIL (b562R


TEV proteas


His(10)


	N-term
	I	L	N	S	S		D	C	P
19


				TM1
K	A	G	R	H	N	Y	I	F	V
29



M	I	P	T	L	Y	S	I	I	F
39



V	V	G	I	F	G	N	S	L	V
49



V	I	V	I	Y	F	Y	M	K	L
59


ICL1				TM2
K	T	V	A	S	V	F	L	L	N
69



L	A	L	A	D	L	C	F	L	L
79



T	L	P	L	W	A	V	Y	T	A
89


		ECL1					TM3
M	E	Y	R	W	P	F	G	N	Y
99



L	C	K	I	A	S	A	S	V	S
109



F	N	L	Y	A	S	V	F	L	L
119



T	C	L	S	I	D	R	Y	L	A
129


			ICL2
I	V	H	P	M	K	S	R	L	R
139


	TM4
R	T	M	L	V	A	K	V	T	C
149



I	I	I	W	L	L	A	G	L	A
159



S	L	P	A	I	I	H	R	N	V
169


ECL2
F	F	I	E	N	T	N	I	T	V
179



C	A	F	H	Y	E	S	Q	N	S
189


	TM5
T	L	P	I	G	L	G	L	T	K
199



N	I	L	G	F	L	F	P	F	L
209



I	I	L	T	S	Y	T	L	I	W
219


						ICL3
K	A	L	K	K	A	Y	E	I	Q
229


			TM6
K	N	K	P	R	N	D	D	I	F
239



K	I	I	M	A	I	V	L	F	F
249



F	F	S	W	I	P	H	Q	I	F
259



T	F	L	D	V	L	I	Q	L	G
269


ECL3				TM7
I	I	R	D	C	R	I	A	D	I
279



V	D	T	A	M	P	I	T	I	C
289



I	A	Y	F	N	N	C	L	N	P
299


						H8
L	F	Y	G	F	L	G	K	K	F
309



K	R	Y	F	L	Q

SCHEMATIC

Perspective: Wild-type Construct

N-term

TM1

ICL1

TM2

ECL1

TM3

ICL2

TM4

ECL2

TM5

ICL3

TM6

ECL3

TM7

H8

C-term

hemagg

FLAG

BRIL (

TEV pr

His(10

N-term

TM1

ICL1

TM2

ECL1

TM3

ICL2

TM4

ECL2

TM5

ICL3

TM6

ECL3

TM7

H8

DIAGRAMS

MUTATIONS

Position	Wild-type	Mutated

DELETIONS

from 1 to 1

from 7 to 16

from 316 to 359

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: fusion

Name: BRIL (b562RIL)

Sequence: None

Deletions:

Positions: N-term_2 None:None

Presence in Crystal:YES

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: tag

Name: His(10)

Sequence: None

Deletions:

Positions: N-term_4 None:None

Presence in Crystal:NO

Remarks: None

EXPRESSION

Expression method	Host cell type	Host cell	Remarks
Baculovirus	Sf9	Insect	Bac-to-Bac

SOLUBILIZATION

Solvent type	Item	Concentration
detergent	DDM	1.0 %w/v
additive	CHS	0.2 %w/v

Remarks:

PURIFICATION

Purification type	Description
Proteolysis by TEV protease	None
Deglycosylation by PNGase F	None

Remarks:None

CRYSTALLIZATION

METHOD: in meso/LCP

TYPE: lipidic cubic phase (LCP) (monoolein (9.9 MAG))

PROTEIN CONCENTRATION: 30 mg/ml

Temperature: 20°C

ph : 5.0-5.0

Remarks:

Chemical lists

Type	Item	Concentration
Additional
unknown	Sodium citrate, pH 5.0	100 mM
unknown	KH2PO4	400 mM
unknown	PEG400	25 %(v/v)
unknown	DMSO	6 %
Detergent
unknown	DDM	0.02 %w/v
Lipid
unknown	CHS	0.004 %w/v