GPCRdb

grm5_human

NAME

5CGC

ORGANISM

Human (Homo sapiens)

PROTEIN NAME

grm5_human

SOURCE

SWISSPROT

SEQUENCE

Perspective: Wild-type Construct


N-term
M	V	L	L	L	I	L	S	V	L
10



L	L	K	E	D	V	R	G	S	A
20



Q	S	S	E	R	R	V	V	A	H
30



M	P	G	D	I	I	I	G	A	L
40



F	S	V	H	H	Q	P	T	V	D
50



K	V	H	E	R	K	C	G	A	V
60



R	E	Q	Y	G	I	Q	R	V	E
70



A	M	L	H	T	L	E	R	I	N
80



S	D	P	T	L	L	P	N	I	T
90



L	G	C	E	I	R	D	S	C	W
100



H	S	A	V	A	L	E	Q	S	I
110



E	F	I	R	D	S	L	I	S	S
120



E	E	E	E	G	L	V	R	C	V
130



D	G	S	S	S	S	F	R	S	K
140



K	P	I	V	G	V	I	G	P	G
150



S	S	S	V	A	I	Q	V	Q	N
160



L	L	Q	L	F	N	I	P	Q	I
170



A	Y	S	A	T	S	M	D	L	S
180



D	K	T	L	F	K	Y	F	M	R
190



V	V	P	S	D	A	Q	Q	A	R
200



A	M	V	D	I	V	K	R	Y	N
210



W	T	Y	V	S	A	V	H	T	E
220



G	N	Y	G	E	S	G	M	E	A
230



F	K	D	M	S	A	K	E	G	I
240



C	I	A	H	S	Y	K	I	Y	S
250



N	A	G	E	Q	S	F	D	K	L
260



L	K	K	L	T	S	H	L	P	K
270



A	R	V	V	A	C	F	C	E	G
280



M	T	V	R	G	L	L	M	A	M
290



R	R	L	G	L	A	G	E	F	L
300



L	L	G	S	D	G	W	A	D	R
310



Y	D	V	T	D	G	Y	Q	R	E
320



A	V	G	G	I	T	I	K	L	Q
330



S	P	D	V	K	W	F	D	D	Y
340



Y	L	K	L	R	P	E	T	N	H
350



R	N	P	W	F	Q	E	F	W	Q
360



H	R	F	Q	C	R	L	E	G	F
370



P	Q	E	N	S	K	Y	N	K	T
380



C	N	S	S	L	T	L	K	T	H
390



H	V	Q	D	S	K	M	G	F	V
400



I	N	A	I	Y	S	M	A	Y	G
410



L	H	N	M	Q	M	S	L	C	P
420



G	Y	A	G	L	C	D	A	M	K
430



P	I	D	G	R	K	L	L	E	S
440



L	M	K	T	N	F	T	G	V	S
450



G	D	T	I	L	F	D	E	N	G
460



D	S	P	G	R	Y	E	I	M	N
470



F	K	E	M	G	K	D	Y	F	D
480



Y	I	N	V	G	S	W	D	N	G
490



E	L	K	M	D	D	D	E	V	W
500



S	K	K	S	N	I	I	R	S	V
510



C	S	E	P	C	E	K	G	Q	I
520



K	V	I	R	K	G	E	V	S	C
530



C	W	T	C	T	P	C	K	E	N
540



E	Y	V	F	D	E	Y	T	C	K
550



A	C	Q	L	G	S	W	P	T	D
560



D	L	T	G	C	D	L	I	P	V
570


							TM1
Q	Y	L	R	W	G	D	P	E	P
580



I	A	A	V	V	F	A	C	L	G
590



L	L	A	T	L	F	V	T	V	V
600


				ICL1
F	I	I	Y	R	D	T	P	V	V
610


			TM2
K	S	S	S	R	E	L	C	Y	I
620



I	L	A	G	I	C	L	G	Y	L
630


						ECL1
C	T	F	C	L	I	A	K	P	K
640


TM3
Q	I	Y	C	Y	L	Q	R	I	G
650



I	G	L	S	P	A	M	S	Y	S
660



A	L	V	T	K	T	N	R	I	A
670


			ICL2
R	I	L	A	G	S	K	K	K	I
680



C	T	K	K	P	R	F	M	S	A
690


TM4
C	A	Q	L	V	I	A	F	I	L
700



I	C	I	Q	L	G	I	I	V	A
710


					ECL2
L	F	I	M	E	P	P	D	I	M
720



H	D	Y	P	S	I	R	E	V	Y
730


					TM5
L	I	C	N	T	T	N	L	G	V
740



V	T	P	L	G	Y	N	G	L	L
750



I	L	S	C	T	F	Y	A	F	K
760


	ICL3
T	R	N	V	P	A	N	F	N	E
770


TM6
A	K	Y	I	A	F	T	M	Y	T
780



T	C	I	I	W	L	A	F	V	P
790


					ECL3				TM7
I	Y	F	G	S	N	Y	K	I	I
800



T	M	C	F	S	V	S	L	S	A
810



T	V	A	L	G	C	M	F	V	P
820


								H8
K	V	Y	I	I	L	A	K	P	E
830



R	N	V	R	S	A	F	T	T	S
840


C-term
T	V	V	R	M	H	V	G	D	G
850



K	S	S	S	A	A	S	R	S	S
860



S	L	V	N	L	W	K	R	R	G
870



S	S	G	E	T	L	R	Y	K	D
880



R	R	L	A	Q	H	K	S	E	I
890



E	C	F	T	P	K	G	S	M	G
900



N	G	G	R	A	T	M	S	S	S
910



N	G	K	S	V	T	W	A	Q	N
920



E	K	S	S	R	G	Q	H	L	W
930



Q	R	L	S	I	H	I	N	K	K
940



E	N	P	N	Q	T	A	V	I	K
950



P	F	P	K	S	T	E	S	R	G
960



L	G	A	G	A	G	A	G	G	S
970



A	G	G	V	G	A	T	G	G	A
980



G	C	A	G	A	G	P	G	G	P
990



E	S	P	D	A	G	P	K	A	L
1000



Y	D	V	A	E	A	E	E	H	F
1010



P	A	P	A	R	P	R	S	P	S
1020



P	I	S	T	L	S	H	R	A	G
1030



S	A	S	R	T	D	D	D	V	P
1040



S	L	H	S	E	P	V	A	R	S
1050



S	S	S	Q	G	S	L	M	E	Q
1060



I	S	S	V	V	T	R	F	T	A
1070



N	I	S	E	L	N	S	M	M	L
1080



S	T	A	A	P	S	P	G	V	G
1090



A	P	L	C	S	S	Y	L	I	P
1100



K	E	I	Q	L	P	T	T	M	T
1110



T	F	A	E	I	Q	P	L	P	A
1120



I	E	V	T	G	G	A	Q	P	A
1130



A	G	A	Q	A	A	G	D	A	A
1140



R	E	S	P	A	A	G	P	E	A
1150



A	A	A	K	P	D	L	E	E	L
1160



V	A	L	T	P	P	S	P	F	R
1170



D	S	V	D	S	G	S	T	T	P
1180



N	S	P	V	S	E	S	A	L	C
1190



I	P	S	S	P	K	Y	D	T	L
1200



I	I	R	D	Y	T	Q	S	S	S
1210



S	L


GP64 signal


	N-term
	P	V	Q	Y	L	R	W	G	D
577


TM1
P	A	P	I	A	A	V	V	F	A
587



C	L	G	L	L	A	T	L	F	V
597



T	V	V	F	I	I	Y	R	D	T
607


ICL1						TM2
P	V	V	K	S	S	S	R	E	L
617



C	Y	I	I	L	A	G	I	C	L
627



G	Y	L	C	T	F	C	L	I	A
637


ECL1				TM3
K	P	K	Q	I	Y	C	Y	L	Q
647



R	I	G	I	G	L	S	P	A	M
657



S	Y	S	A	L	V	T	K	T	Y
667


						ICL2
R	A	A	R	I	L	A	M	S	K
677



K	K	I	C	T	K	K	P	R
686


		TM4
F	M	S	A	C	A	Q	L	V	I
696



A	F	I	L	I	C	I	Q	L	G
706



I	I	V	A	L	F	I	M	E	P
716


ECL2
P	D	I	M	H	D	Y	P	S	I
726



R	E	V	Y	L	I	C	N	T	T
736


TM5
N	L	G	V	V	A	P	L	G	Y
746



N	G	L	L	I	L	A	C	T	F
756


					ICL3
Y	A	F	K	T	R	N	V	P	A
766


			TM6
N	F	N	E	A	K	Y	I	A	F
776



T	M	Y	T	T	C	I	I	W	L
786



A	F	V	P	I	Y	F	G	S	N
796


TM7
Y	K	I	I	T	M	C	F	S	V
806



S	L	S	A	T	V	A	L	G	C
816



M	F	V	P	K	V	Y	I	I	L
826

H8

A

K

P

E

R

N

V

R

S

A

836


His(10)

SCHEMATIC

Perspective: Wild-type Construct

N-term

TM1

ICL1

TM2

ECL1

TM3

ICL2

TM4

ECL2

TM5

ICL3

TM6

ECL3

TM7

H8

C-term

GP64 s

N-term

TM1

ICL1

TM2

ECL1

TM3

T4 Lys

ICL2

TM4

ECL2

TM5

ICL3

TM6

ECL3

TM7

H8

His(10

DIAGRAMS

MUTATIONS

Position	Wild-type	Mutated
579 \|	E	A
667 \|	N	Y
669 \|	I	A
675 \|	G	M
742 \|	T	A
753 \|	S	A

DELETIONS

from 1 to 568

from 837 to 1212

MODIFICATIONS

AUXILIARY PROTEINS

Protein_type: signal

Name: GP64 signal

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_1 678:679

Presence in Crystal:YES

Remarks: None

Protein_type: tag

Name: His(10)

Sequence: None

Deletions:

Positions: C-term_2 None:None

Presence in Crystal:YES

Remarks: None

EXPRESSION

Expression method	Host cell type	Host cell	Remarks
Baculovirus	Sf21	Insect	FastBac

SOLUBILIZATION

Solvent type	Item	Concentration
detergent	DDM	1.5 %w/v

Remarks:

PURIFICATION

Purification type	Description
Alkylation by Iodoacetamide	None

Remarks:None

CRYSTALLIZATION

METHOD: in meso/LCP

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

PROTEIN CONCENTRATION: 40 mg/ml

Temperature: 20°C

ph : 5.5-6.8

Remarks: The final protein:lipid ratio was 40:60 (w/w)

Chemical lists

Type	Item	Concentration
Additional
unknown	MES, pH 5.5-6.8	100 mM
unknown	(NH4)2HPO4	100-250 mM
unknown	PEG400	24-34 %(v/v)
Detergent
detergent	DDM	0.03 % w/v