GPCRdb

ntr1_rat

NAME

5T04

ORGANISM

Rat (Rattus norvegicus)

PROTEIN NAME

ntr1_rat

SOURCE

SWISSPROT

SEQUENCE

Perspective: Wild-type Construct


N-term
M	H	L	N	S	S	V	P	Q	G
10



T	P	G	E	P	D	A	Q	P	F
20



S	G	P	Q	S	E	M	E	A	T
30



F	L	A	L	S	L	S	N	G	S
40



G	N	T	S	E	S	D	T	A	G
50



P	N	S	D	L	D	V	N	T	D
60


TM1
I	Y	S	K	V	L	V	T	A	I
70



Y	L	A	L	F	V	V	G	T	V
80



G	N	S	V	T	A	F	T	L	A
90


		ICL1				TM2
R	K	K	S	L	Q	S	L	Q	S
100



T	V	H	Y	H	L	G	S	L	A
110



L	S	D	L	L	I	L	L	L	A
120



M	P	V	E	L	Y	N	F	I	W
130


	ECL1						TM3
V	H	H	P	W	A	F	G	D	A
140



G	C	R	G	Y	Y	F	L	R	D
150



A	C	T	Y	A	T	A	L	N	V
160



A	S	L	S	V	E	R	Y	L	A
170


			ICL2
I	C	H	P	F	K	A	K	T	L
180


	TM4
M	S	R	S	R	T	K	K	F	I
190



S	A	I	W	L	A	S	A	L	L
200



A	I	P	M	L	F	T	M	G	L
210


ECL2
Q	N	R	S	G	D	G	T	H	P
220



G	G	L	V	C	T	P	I	V	D
230


TM5
T	A	T	V	K	V	V	I	Q	V
240



N	T	F	M	S	F	L	F	P	M
250



L	V	I	S	I	L	N	T	V	I
260



A	N	K	L	T	V	M	V	H	Q
270


					ICL3
A	A	E	Q	G	R	V	C	T	V
280



G	T	H	N	G	L	E	H	S	T
290


						TM6
F	N	M	T	I	E	P	G	R	V
300



Q	A	L	R	H	G	V	L	V	L
310



R	A	V	V	I	A	F	V	V	C
320



W	L	P	Y	H	V	R	R	L	M
330


				ECL3
F	C	Y	I	S	D	E	Q	W	T
340


TM7
T	F	L	F	D	F	Y	H	Y	F
350



Y	M	L	T	N	A	L	F	Y	V
360



S	S	A	I	N	P	I	L	Y	N
370


		H8
L	V	S	A	N	F	R	Q	V	F
380



L	S	T	L	A	C	L	C	P	G
390


C-term
W	R	H	R	R	K	K	R	P	T
400



F	S	R	K	P	N	S	M	S	S
410



N	H	A	F	S	T	S	A	T	R
420



E	T	L	Y


hemagglutin


FLAG


	N-term
	T	S	E	S	D	T	A	G	P
51



N	S	D	L	D	V	N	T	D	I
61


TM1
Y	S	K	V	L	V	T	A	I	Y
71



L	A	L	F	V	V	G	T	V	G
81



N	S	V	T	L	F	T	L	A	R
91


	ICL1				TM2
K	K	S	L	Q	S	L	Q	S	T
101



V	H	Y	H	L	G	S	L	A	L
111



S	D	L	L	I	L	L	L	A	M
121



P	V	E	L	Y	N	F	I	W	V
131


ECL1						TM3
H	H	P	W	A	F	G	D	A	G
141



C	R	G	Y	Y	F	L	R	D	A
151



C	T	Y	A	T	A	L	N	V	A
161



S	L	S	V	E	R	Y	L	A	I
171


		ICL2
C	H	P	F	K	A	K	T	L	M
181


TM4
S	R	S	R	T	K	K	F	I	S
191



A	I	W	L	A	S	A	L	L	A
201



I	P	M	L	F	T	M	G	L	Q
211


ECL2
N	R	S	A	D	G	T	H	P	G
221



G	L	V	C	T	P	I	V	D	T
231


TM5
A	T	V	K	V	V	I	Q	V	N
241



T	F	M	S	F	L	F	P	M	L
251



V	I	S	I	L	N	T	V	I	A
261



N	K	L	T	V	M	V


T4 Lysozyme


TM6
P	G	R	V	Q	A	L	R
304



H	G	V	L	V	L	R	A	V	V
314



I	A	F	V	V	C	W	L	P	Y
324



H	V	R	R	L	M	F	C	Y	I
334


ECL3					TM7
S	D	E	Q	W	T	T	F	L	F
344



D	F	Y	H	Y	F	Y	M	L	T
354



N	A	L	A	Y	A	S	S	A	I
364


								H8
N	P	I	L	Y	N	L	V	S	A
374



N	F	R	Q	V	F	L	S	T	L
384


				C-term
A	C	L	C	P	G	W	R	H	R
394



R	K


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

hemagg

FLAG

N-term

TM1

ICL1

TM2

ECL1

TM3

ICL2

TM4

ECL2

TM5

T4 Lys

TM6

ECL3

TM7

H8

C-term

His(10

DIAGRAMS

MUTATIONS

Position	Wild-type	Mutated
86 \|	A	L
215 \|	G	A
358 \|	F	A
360 \|	V	A

DELETIONS

from 1 to 42

from 269 to 296

from 397 to 424

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:YES

Remarks: None

Protein_type: fusion

Name: T4 Lysozyme (T4L)

Sequence: None

Deletions:

Positions: Within Receptor_2 269:296

Presence in Crystal:YES

Remarks: None

Protein_type: linker

Name: GSGS

Sequence: None

Deletions:

Positions: Within Receptor_3 296:296

Presence in Crystal:YES

Remarks: None

Protein_type: tag

Name: His(10)

Sequence: None

Deletions:

Positions: C-term_4 None:None

Presence in Crystal:YES

Remarks: None

EXPRESSION

Expression method	Host cell type	Host cell	Remarks
Baculovirus	Tni PRO (Trichoplusia ni) cell lines	Insect

SOLUBILIZATION

Solvent type	Item	Concentration
detergent	LMNG	3.0 %w/v
additive	CHS	0.3 %w/v

Remarks:

PURIFICATION

Purification type	Description
NONE	None

Remarks:None

CRYSTALLIZATION

METHOD: in meso/LCP

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

PROTEIN CONCENTRATION: 60 mg/ml

Temperature: 20°C

ph : 8.5-9.0

Remarks:

Chemical lists

Type	Item	Concentration
Detergent
detergent	LMNG	0.003 %w/v
Lipid
lipid	CHS	0.0003 %w/v
Precipitant
unknown	PEG 400	13–16 %v/v
Buffer
unknown	TrisHCl pH 8.5–9.0	80.0 mM
Additional
unknown	TCEP	1.9 mM
unknown	lithium acetate	68-91 mM