PDB-3cus: Structure of a double ILE/PHE mutant of NI-FE hydrogenase refined...

Basic information

• Entry: Database: PDB / ID: 3cus
• Title: Structure of a double ILE/PHE mutant of NI-FE hydrogenase refined at 2.2 angstrom resolution
• Components: (Periplasmic [NiFe] hydrogenase ...) x 2
• Keywords: OXIDOREDUCTASE / NI-FE hydrogenase tunnel mutant / Iron / Iron-sulfur / Metal-binding / Nickel

Function / homology

Function:

cytochrome-c3 hydrogenase / cytochrome-c3 hydrogenase activity / ferredoxin hydrogenase complex / [Ni-Fe] hydrogenase complex / ferredoxin hydrogenase activity / anaerobic respiration / 3 iron, 4 sulfur cluster binding / nickel cation binding / 4 iron, 4 sulfur cluster binding / periplasmic space / electron transfer activity / metal ion binding / membrane

Domain/homology:

Cytochrome-c3 Hydrogenase; Chain A, domain 2 / Cytochrome-c3 hydrogenase, C-terminal domain / NADH:ubiquinone oxidoreductase-like, 20kDa subunit / : / [NiFe]-hydrogenase, small subunit / Cytochrome-c3 hydrogenase, C-terminal / [NiFe]-hydrogenase, small subunit, C-terminal domain superfamily / NiFe/NiFeSe hydrogenase small subunit C-terminal / Nickel-dependent hydrogenases large subunit signature 1. / Nickel-dependent hydrogenases large subunit signature 2. / [NiFe]-hydrogenase, small subunit, N-terminal domain superfamily / Nickel-dependent hydrogenase, large subunit, nickel binding site / Nickel-dependent hydrogenase, large subunit / Nickel-dependent hydrogenase / Cytochrome-c3 Hydrogenase; chain B / Cytochrome-c3 Hydrogenase, chain B / Twin-arginine translocation pathway, signal sequence, bacterial/archaeal / NADH:ubiquinone oxidoreductase-like, 20kDa subunit / NADH ubiquinone oxidoreductase, 20 Kd subunit / [NiFe]-hydrogenase, large subunit / Twin arginine translocation (Tat) signal profile. / Twin-arginine translocation pathway, signal sequence / Few Secondary Structures / Irregular / Rossmann fold / Orthogonal Bundle / 3-Layer(aba) Sandwich / Mainly Alpha / Alpha Beta

Component:

FE3-S4 CLUSTER / CARBONMONOXIDE-(DICYANO) IRON / NICKEL (II) ION / IRON/SULFUR CLUSTER / Periplasmic [NiFe] hydrogenase small subunit / Periplasmic [NiFe] hydrogenase large subunit

• Biological species: Desulfovibrio fructosovorans (bacteria)
• Method: X-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.2 Å
• Authors: Volbeda, A.

Citation

Journal: Proc.Natl.Acad.Sci.Usa / Year: 2008
Title: Experimental approaches to kinetics of gas diffusion in hydrogenase
Authors: Leroux, F. / Dementin, S. / Burlat, B. / Cournac, L. / Volbeda, A. / Champ, S. / Martin, L. / Guigliarelli, B. / Bertrand, P. / Fontecilla-Camps, J. / Rousset, M.

#1: Journal: Nat.Struct.Biol. / Year: 1997
Title: Gas access to the active site of Ni-Fe hydrogenases probed by X-ray crystallography and molecular dynamics
Authors: Montet, Y. / Amara, P. / Volbeda, A. / Vernede, X. / Hatchikian, E.C. / Field, M.J. / Frey, M. / Fontecilla-Camps, J.C.

#2: Journal: Int.J.Hydrogen Energy / Year: 2002
Title: High-Resolution Crystallographic Analysis of Desulfovibrio Fructosovorans [Nife] Hydrogenase
Authors: Volbeda, A. / Montet, Y. / Vernede, X. / Hatchikian, E.C. / Fontecilla-Camps, J.C.

#3: Journal: J.Biol.Inorg.Chem. / Year: 2005
Title: Structural differences between the ready and unready oxidized states of [NiFe] hydrogenases
Authors: Volbeda, A. / Martin, L. / Cavazza, C. / Matho, M. / Faber, B.W. / Roseboom, W. / Albracht, S.P. / Garcin, E. / Rousset, M. / Fontecilla-Camps, J.C.

History

Deposition	Apr 17, 2008	Deposition site: RCSB / Processing site: PDBJ
Revision 1.0	Aug 5, 2008	Provider: repository / Type: Initial release
Revision 1.1	Jul 13, 2011	Group: Non-polymer description / Version format compliance
Revision 1.2	Sep 7, 2011	Group: Database references
Revision 1.3	Nov 1, 2023	Group: Data collection / Database references / Derived calculations / Refinement description Category: chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / pdbx_struct_conn_angle / struct_conn / struct_ncs_dom_lim / struct_ref_seq_dif / struct_site Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_conn_angle.ptnr1_auth_asym_id / _pdbx_struct_conn_angle.ptnr1_auth_comp_id / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_asym_id / _pdbx_struct_conn_angle.ptnr1_label_atom_id / _pdbx_struct_conn_angle.ptnr1_label_comp_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr2_auth_asym_id / _pdbx_struct_conn_angle.ptnr2_auth_comp_id / _pdbx_struct_conn_angle.ptnr2_auth_seq_id / _pdbx_struct_conn_angle.ptnr2_label_asym_id / _pdbx_struct_conn_angle.ptnr2_label_atom_id / _pdbx_struct_conn_angle.ptnr2_label_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_asym_id / _pdbx_struct_conn_angle.ptnr3_auth_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_asym_id / _pdbx_struct_conn_angle.ptnr3_label_atom_id / _pdbx_struct_conn_angle.ptnr3_label_comp_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _struct_conn.pdbx_dist_value / _struct_conn.pdbx_leaving_atom_flag / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_comp_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_atom_id / _struct_conn.ptnr1_label_comp_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_auth_comp_id / _struct_conn.ptnr2_auth_seq_id / _struct_conn.ptnr2_label_asym_id / _struct_conn.ptnr2_label_atom_id / _struct_conn.ptnr2_label_comp_id / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ncs_dom_lim.end_label_asym_id / _struct_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Revision 1.4	Nov 15, 2023	Group: Data collection / Category: chem_comp_atom / chem_comp_bond / Item: _chem_comp_atom.atom_id / _chem_comp_bond.atom_id_2
Revision 1.5	Nov 20, 2024	Group: Structure summary / Category: pdbx_entry_details / pdbx_modification_feature

Assembly

Deposited unit

A: Periplasmic [NiFe] hydrogenase small subunit

Q: Periplasmic [NiFe] hydrogenase large subunit

B: Periplasmic [NiFe] hydrogenase small subunit

R: Periplasmic [NiFe] hydrogenase large subunit

C: Periplasmic [NiFe] hydrogenase small subunit

S: Periplasmic [NiFe] hydrogenase large subunit

hetero molecules

Summary:

• 269 kDa, 6 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	268,665	29
Polymers	264,551	6
Non-polymers	4,114	23
Water	18,249	1013

1

A: Periplasmic [NiFe] hydrogenase small subunit

Q: Periplasmic [NiFe] hydrogenase large subunit

hetero molecules

Summary:

• defined by author&software
• 89.7 kDa, 2 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	89,678	11
Polymers	88,184	2
Non-polymers	1,494	9
Water	36	2

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

Calculated values:

Buried area	9220 Å2
ΔGint	-122.5 kcal/mol
Surface area	25290 Å2
Method	PISA

2

B: Periplasmic [NiFe] hydrogenase small subunit

R: Periplasmic [NiFe] hydrogenase large subunit

hetero molecules

Summary:

• defined by author&software
• 89.5 kDa, 2 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	89,494	9
Polymers	88,184	2
Non-polymers	1,310	7
Water	36	2

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

Calculated values:

Buried area	9020 Å2
ΔGint	-121.2 kcal/mol
Surface area	25350 Å2
Method	PISA

3

C: Periplasmic [NiFe] hydrogenase small subunit

S: Periplasmic [NiFe] hydrogenase large subunit

hetero molecules

Summary:

• defined by author&software
• 89.5 kDa, 2 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	89,494	9
Polymers	88,184	2
Non-polymers	1,310	7
Water	36	2

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

Calculated values:

Buried area	8820 Å2
ΔGint	-121.2 kcal/mol
Surface area	25020 Å2
Method	PISA

Unit cell

Length a, b, c (Å)	62.400, 99.960, 182.690
Angle α, β, γ (deg.)	90.00, 92.23, 90.00
Int Tables number	4
Space group name H-M	P1211

Noncrystallographic symmetry (NCS)

NCS domain:

ID	Ens-ID	Details
1	1	A
2	1	B
3	1	C
4	1	A
5	1	B
6	1	C
7	1	A
8	1	B
9	1	C
10	1	A
11	1	B
12	1	C
13	1	A
14	1	B
15	1	C
16	1	A
17	1	B
18	1	C
19	1	A
20	1	B
21	1	C
22	1	A
23	1	B
24	1	C
25	1	A
26	1	B
27	1	C
28	1	A
29	1	B
30	1	C
31	1	A
32	1	B
33	1	C
34	1	A
35	1	B
36	1	C
37	1	A
38	1	B
39	1	C
40	1	A
41	1	B
42	1	C
43	1	A
44	1	B
45	1	C
46	1	A
47	1	B
48	1	C
49	1	A
50	1	B
51	1	C
52	1	A
53	1	B
54	1	C
55	1	A
56	1	B
57	1	C
58	1	A
59	1	B
60	1	C
61	1	A
62	1	B
63	1	C
64	1	A
65	1	B
66	1	C
67	1	A
68	1	B
69	1	C
70	1	A
71	1	B
72	1	C
73	1	A
74	1	B
75	1	C
76	1	A
77	1	B
78	1	C
79	1	A
80	1	B
81	1	C
82	1	A
83	1	B
84	1	C
85	1	A
86	1	B
87	1	C
88	1	A
89	1	B
90	1	C
91	1	A
92	1	B
93	1	C
94	1	A
95	1	B
96	1	C
97	1	A
98	1	B
99	1	C
100	1	A
101	1	B
102	1	C
103	1	A
104	1	B
105	1	C
106	1	A
107	1	B
108	1	C
109	1	A
110	1	B
111	1	C
112	1	A
113	1	B
114	1	C
1	2	A
2	2	B
3	2	C
4	2	A
5	2	B
6	2	C
7	2	A
8	2	B
9	2	C
10	2	A
11	2	B
12	2	C
13	2	A
14	2	B
15	2	C
16	2	A
17	2	B
18	2	C
19	2	A
20	2	B
21	2	C
22	2	A
23	2	B
24	2	C
25	2	A
26	2	B
27	2	C
28	2	A
29	2	B
30	2	C
31	2	A
32	2	B
33	2	C
34	2	A
35	2	B
36	2	C
37	2	A
38	2	B
39	2	C
40	2	A
41	2	B
42	2	C
43	2	A
44	2	B
45	2	C
46	2	A
47	2	B
48	2	C
49	2	A
50	2	B
51	2	C
52	2	A
53	2	B
54	2	C
55	2	A
56	2	B
57	2	C
58	2	A
59	2	B
60	2	C
61	2	A
62	2	B
63	2	C
64	2	A
65	2	B
66	2	C
67	2	A
68	2	B
69	2	C
70	2	A
71	2	B
72	2	C
73	2	A
74	2	B
75	2	C
76	2	A
77	2	B
78	2	C
79	2	A
80	2	B
81	2	C
82	2	A
83	2	B
84	2	C
85	2	A
86	2	B
87	2	C
1	3	Q
2	3	R
3	3	S
4	3	Q
5	3	R
6	3	S
7	3	Q
8	3	R
9	3	S
10	3	Q
11	3	R
12	3	S
13	3	Q
14	3	R
15	3	S
16	3	Q
17	3	R
18	3	S
19	3	Q
20	3	R
21	3	S
22	3	Q
23	3	R
24	3	S
25	3	Q
26	3	R
27	3	S
28	3	Q
29	3	R
30	3	S
31	3	Q
32	3	R
33	3	S
34	3	Q
35	3	R
36	3	S
37	3	Q
38	3	R
39	3	S
40	3	Q
41	3	R
42	3	S
43	3	Q
44	3	R
45	3	S
46	3	Q
47	3	R
48	3	S
49	3	Q
50	3	R
51	3	S
52	3	Q
53	3	R
54	3	S
55	3	Q
56	3	R
57	3	S
58	3	Q
59	3	R
60	3	S
61	3	Q
62	3	R
63	3	S
64	3	Q
65	3	R
66	3	S
67	3	Q
68	3	R
69	3	S
70	3	Q
71	3	R
72	3	S
73	3	Q
74	3	R
75	3	S
76	3	Q
77	3	R
78	3	S
79	3	Q
80	3	R
81	3	S
82	3	Q
83	3	R
84	3	S
85	3	Q
86	3	R
87	3	S
88	3	Q
89	3	R
90	3	S
91	3	Q
92	3	R
93	3	S
94	3	Q
95	3	R
96	3	S
97	3	Q
98	3	R
99	3	S
100	3	Q
101	3	R
102	3	S
103	3	Q
104	3	R
105	3	S
106	3	Q
107	3	R
108	3	S
1	4	Q
2	4	R
3	4	S
4	4	Q
5	4	R
6	4	S
7	4	Q
8	4	R
9	4	S
10	4	Q
11	4	R
12	4	S
13	4	Q
14	4	R
15	4	S
16	4	Q
17	4	R
18	4	S
19	4	Q
20	4	R
21	4	S
22	4	Q
23	4	R
24	4	S
25	4	Q
26	4	R
27	4	S
28	4	Q
29	4	R
30	4	S
31	4	Q
32	4	R
33	4	S
34	4	Q
35	4	R
36	4	S
37	4	Q
38	4	R
39	4	S
40	4	Q
41	4	R
42	4	S
43	4	Q
44	4	R
45	4	S
46	4	Q
47	4	R
48	4	S
49	4	Q
50	4	R
51	4	S
52	4	Q
53	4	R
54	4	S
55	4	Q
56	4	R
57	4	S
58	4	Q
59	4	R
60	4	S
61	4	Q
62	4	R
63	4	S
64	4	Q
65	4	R
66	4	S

NCS domain segments:

Dom-ID	Component-ID	Ens-ID	Beg auth comp-ID	Beg label comp-ID	End auth comp-ID	End label comp-ID	Refine code	Auth asym-ID	Label asym-ID	Auth seq-ID	Label seq-ID
1	1	1	PRO	PRO	PRO	PRO	3	A	A	7	7
2	1	1	PRO	PRO	PRO	PRO	3	B	C	7	7
3	1	1	PRO	PRO	PRO	PRO	3	C	E	7	7
4	2	1	SER	SER	ILE	ILE	1	A	A	8 - 28	8 - 28
5	2	1	SER	SER	ILE	ILE	1	B	C	8 - 28	8 - 28
6	2	1	SER	SER	ILE	ILE	1	C	E	8 - 28	8 - 28
7	3	1	LYS	LYS	PRO	PRO	3	A	A	29 - 30	29 - 30
8	3	1	LYS	LYS	PRO	PRO	3	B	C	29 - 30	29 - 30
9	3	1	LYS	LYS	PRO	PRO	3	C	E	29 - 30	29 - 30
10	4	1	TYR	TYR	ILE	ILE	1	A	A	31 - 36	31 - 36
11	4	1	TYR	TYR	ILE	ILE	1	B	C	31 - 36	31 - 36
12	4	1	TYR	TYR	ILE	ILE	1	C	E	31 - 36	31 - 36
13	5	1	LEU	LEU	THR	THR	3	A	A	37 - 39	37 - 39
14	5	1	LEU	LEU	THR	THR	3	B	C	37 - 39	37 - 39
15	5	1	LEU	LEU	THR	THR	3	C	E	37 - 39	37 - 39
16	6	1	ILE	ILE	GLY	GLY	1	A	A	40 - 53	40 - 53
17	6	1	ILE	ILE	GLY	GLY	1	B	C	40 - 53	40 - 53
18	6	1	ILE	ILE	GLY	GLY	1	C	E	40 - 53	40 - 53
19	7	1	GLU	GLU	GLU	GLU	3	A	A	54	54
20	7	1	GLU	GLU	GLU	GLU	3	B	C	54	54
21	7	1	GLU	GLU	GLU	GLU	3	C	E	54	54
22	8	1	ALA	ALA	ALA	ALA	4	A	A	55	55
23	8	1	ALA	ALA	ALA	ALA	4	B	C	55	55
24	8	1	ALA	ALA	ALA	ALA	4	C	E	55	55
25	9	1	ALA	ALA	GLU	GLU	1	A	A	56 - 57	56 - 57
26	9	1	ALA	ALA	GLU	GLU	1	B	C	56 - 57	56 - 57
27	9	1	ALA	ALA	GLU	GLU	1	C	E	56 - 57	56 - 57
28	10	1	ALA	ALA	ALA	ALA	4	A	A	58	58
29	10	1	ALA	ALA	ALA	ALA	4	B	C	58	58
30	10	1	ALA	ALA	ALA	ALA	4	C	E	58	58
31	11	1	ALA	ALA	LEU	LEU	1	A	A	59 - 60	59 - 60
32	11	1	ALA	ALA	LEU	LEU	1	B	C	59 - 60	59 - 60
33	11	1	ALA	ALA	LEU	LEU	1	C	E	59 - 60	59 - 60
34	12	1	GLN	GLN	GLN	GLN	2	A	A	62	62
35	12	1	GLN	GLN	GLN	GLN	2	B	C	62	62
36	12	1	GLN	GLN	GLN	GLN	2	C	E	62	62
37	13	1	ALA	ALA	LEU	LEU	1	A	A	63 - 64	63 - 64
38	13	1	ALA	ALA	LEU	LEU	1	B	C	63 - 64	63 - 64
39	13	1	ALA	ALA	LEU	LEU	1	C	E	63 - 64	63 - 64
40	14	1	GLU	GLU	ASP	ASP	3	A	A	65 - 68	65 - 68
41	14	1	GLU	GLU	ASP	ASP	3	B	C	65 - 68	65 - 68
42	14	1	GLU	GLU	ASP	ASP	3	C	E	65 - 68	65 - 68
43	15	1	GLY	GLY	TYR	TYR	1	A	A	69 - 70	69 - 70
44	15	1	GLY	GLY	TYR	TYR	1	B	C	69 - 70	69 - 70
45	15	1	GLY	GLY	TYR	TYR	1	C	E	69 - 70	69 - 70
46	16	1	TYR	TYR	TYR	TYR	2	A	A	71	71
47	16	1	TYR	TYR	TYR	TYR	2	B	C	71	71
48	16	1	TYR	TYR	TYR	TYR	2	C	E	71	71
49	17	1	LEU	LEU	ILE	ILE	1	A	A	72 - 81	72 - 81
50	17	1	LEU	LEU	ILE	ILE	1	B	C	72 - 81	72 - 81
51	17	1	LEU	LEU	ILE	ILE	1	C	E	72 - 81	72 - 81
52	18	1	ASP	ASP	GLN	GLN	3	A	A	82 - 85	82 - 85
53	18	1	ASP	ASP	GLN	GLN	3	B	C	82 - 85	82 - 85
54	18	1	ASP	ASP	GLN	GLN	3	C	E	82 - 85	82 - 85
55	19	1	TRP	TRP	GLY	GLY	1	A	A	86 - 91	86 - 91
56	19	1	TRP	TRP	GLY	GLY	1	B	C	86 - 91	86 - 91
57	19	1	TRP	TRP	GLY	GLY	1	C	E	86 - 91	86 - 91
58	20	1	HIS	HIS	HIS	HIS	3	A	A	92	92
59	20	1	HIS	HIS	HIS	HIS	3	B	C	92	92
60	20	1	HIS	HIS	HIS	HIS	3	C	E	92	92
61	21	1	PRO	PRO	MET	MET	1	A	A	93 - 94	93 - 94
62	21	1	PRO	PRO	MET	MET	1	B	C	93 - 94	93 - 94
63	21	1	PRO	PRO	MET	MET	1	C	E	93 - 94	93 - 94
64	22	1	ILE	ILE	LYS	LYS	3	A	A	95 - 99	95 - 99
65	22	1	ILE	ILE	LYS	LYS	3	B	C	95 - 99	95 - 99
66	22	1	ILE	ILE	LYS	LYS	3	C	E	95 - 99	95 - 99
67	23	1	ALA	ALA	ALA	ALA	1	A	A	101	101
68	23	1	ALA	ALA	ALA	ALA	1	B	C	101	101
69	23	1	ALA	ALA	ALA	ALA	1	C	E	101	101
70	24	1	LYS	LYS	LYS	LYS	3	A	A	104	104
71	24	1	LYS	LYS	LYS	LYS	3	B	C	104	104
72	24	1	LYS	LYS	LYS	LYS	3	C	E	104	104
73	25	1	ALA	ALA	ALA	ALA	4	A	A	105	105
74	25	1	ALA	ALA	ALA	ALA	4	B	C	105	105
75	25	1	ALA	ALA	ALA	ALA	4	C	E	105	105
76	26	1	LYS	LYS	LYS	LYS	3	A	A	106	106
77	26	1	LYS	LYS	LYS	LYS	3	B	C	106	106
78	26	1	LYS	LYS	LYS	LYS	3	C	E	106	106
79	27	1	GLY	GLY	ALA	ALA	1	A	A	107 - 130	107 - 130
80	27	1	GLY	GLY	ALA	ALA	1	B	C	107 - 130	107 - 130
81	27	1	GLY	GLY	ALA	ALA	1	C	E	107 - 130	107 - 130
82	28	1	LYS	LYS	LYS	LYS	2	A	A	131	131
83	28	1	LYS	LYS	LYS	LYS	2	B	C	131	131
84	28	1	LYS	LYS	LYS	LYS	2	C	E	131	131
85	29	1	GLY	GLY	VAL	VAL	1	A	A	132 - 133	132 - 133
86	29	1	GLY	GLY	VAL	VAL	1	B	C	132 - 133	132 - 133
87	29	1	GLY	GLY	VAL	VAL	1	C	E	132 - 133	132 - 133
88	30	1	THR	THR	VAL	VAL	1	A	A	141 - 159	141 - 159
89	30	1	THR	THR	VAL	VAL	1	B	C	141 - 159	141 - 159
90	30	1	THR	THR	VAL	VAL	1	C	E	141 - 159	141 - 159
91	31	1	HIS	HIS	HIS	HIS	2	A	A	160	160
92	31	1	HIS	HIS	HIS	HIS	2	B	C	160	160
93	31	1	HIS	HIS	HIS	HIS	2	C	E	160	160
94	32	1	VAL	VAL	LEU	LEU	3	A	A	161 - 162	161 - 162
95	32	1	VAL	VAL	LEU	LEU	3	B	C	161 - 162	161 - 162
96	32	1	VAL	VAL	LEU	LEU	3	C	E	161 - 162	161 - 162
97	33	1	THR	THR	THR	THR	6	A	A	163	163
98	33	1	THR	THR	THR	THR	6	B	C	163	163
99	33	1	THR	THR	THR	THR	6	C	E	163	163
100	34	1	LYS	LYS	LYS	LYS	3	A	A	164	164
101	34	1	LYS	LYS	LYS	LYS	3	B	C	164	164
102	34	1	LYS	LYS	LYS	LYS	3	C	E	164	164
103	35	1	GLY	GLY	PRO	PRO	2	A	A	165 - 167	165 - 167
104	35	1	GLY	GLY	PRO	PRO	2	B	C	165 - 167	165 - 167
105	35	1	GLY	GLY	PRO	PRO	2	C	E	165 - 167	165 - 167
106	36	1	ASP	ASP	LEU	LEU	3	A	A	168 - 169	168 - 169
107	36	1	ASP	ASP	LEU	LEU	3	B	C	168 - 169	168 - 169
108	36	1	ASP	ASP	LEU	LEU	3	C	E	168 - 169	168 - 169
109	37	1	ASP	ASP	GLU	GLU	6	A	A	170 - 171	170 - 171
110	37	1	ASP	ASP	GLU	GLU	6	B	C	170 - 171	170 - 171
111	37	1	ASP	ASP	GLU	GLU	6	C	E	170 - 171	170 - 171
112	38	1	SF4	SF4	SF4	SF4	1	A	I	267
113	38	1	SF4	SF4	SF4	SF4	1	B	R	267
114	38	1	SF4	SF4	SF4	SF4	1	C	Y	267
1	1	2	GLY	GLY	PRO	PRO	1	A	A	173 - 175	173 - 175
2	1	2	GLY	GLY	PRO	PRO	1	B	C	173 - 175	173 - 175
3	1	2	GLY	GLY	PRO	PRO	1	C	E	173 - 175	173 - 175
4	2	2	LYS	LYS	LEU	LEU	3	A	A	176 - 177	176 - 177
5	2	2	LYS	LYS	LEU	LEU	3	B	C	176 - 177	176 - 177
6	2	2	LYS	LYS	LEU	LEU	3	C	E	176 - 177	176 - 177
7	3	2	PHE	PHE	TYR	TYR	1	A	A	178 - 179	178 - 179
8	3	2	PHE	PHE	TYR	TYR	1	B	C	178 - 179	178 - 179
9	3	2	PHE	PHE	TYR	TYR	1	C	E	178 - 179	178 - 179
10	4	2	GLY	GLY	LEU	LEU	3	A	A	180 - 182	180 - 182
11	4	2	GLY	GLY	LEU	LEU	3	B	C	180 - 182	180 - 182
12	4	2	GLY	GLY	LEU	LEU	3	C	E	180 - 182	180 - 182
13	5	2	VAL	VAL	CYS	CYS	1	A	A	183 - 187	183 - 187
14	5	2	VAL	VAL	CYS	CYS	1	B	C	183 - 187	183 - 187
15	5	2	VAL	VAL	CYS	CYS	1	C	E	183 - 187	183 - 187
16	6	2	PRO	PRO	LEU	LEU	2	A	A	188 - 190	188 - 190
17	6	2	PRO	PRO	LEU	LEU	2	B	C	188 - 190	188 - 190
18	6	2	PRO	PRO	LEU	LEU	2	C	E	188 - 190	188 - 190
19	7	2	PRO	PRO	GLU	GLU	3	A	A	191 - 194	191 - 194
20	7	2	PRO	PRO	GLU	GLU	3	B	C	191 - 194	191 - 194
21	7	2	PRO	PRO	GLU	GLU	3	C	E	191 - 194	191 - 194
22	8	2	ALA	ALA	SER	SER	2	A	A	195 - 196	195 - 196
23	8	2	ALA	ALA	SER	SER	2	B	C	195 - 196	195 - 196
24	8	2	ALA	ALA	SER	SER	2	C	E	195 - 196	195 - 196
25	9	2	GLU	GLU	PHE	PHE	3	A	A	197 - 198	197 - 198
26	9	2	GLU	GLU	PHE	PHE	3	B	C	197 - 198	197 - 198
27	9	2	GLU	GLU	PHE	PHE	3	C	E	197 - 198	197 - 198
28	10	2	ALA	ALA	PHE	PHE	1	A	A	199 - 202	199 - 202
29	10	2	ALA	ALA	PHE	PHE	1	B	C	199 - 202	199 - 202
30	10	2	ALA	ALA	PHE	PHE	1	C	E	199 - 202	199 - 202
31	11	2	ASP	ASP	ASP	ASP	3	A	A	203	203
32	11	2	ASP	ASP	ASP	ASP	3	B	C	203	203
33	11	2	ASP	ASP	ASP	ASP	3	C	E	203	203
34	12	2	SER	SER	SER	SER	1	A	A	204	204
35	12	2	SER	SER	SER	SER	1	B	C	204	204
36	12	2	SER	SER	SER	SER	1	C	E	204	204
37	13	2	GLU	GLU	GLU	GLU	3	A	A	205 - 206	205 - 206
38	13	2	GLU	GLU	GLU	GLU	3	B	C	205 - 206	205 - 206
39	13	2	GLU	GLU	GLU	GLU	3	C	E	205 - 206	205 - 206
40	14	2	ALA	ALA	LYS	LYS	1	A	A	207 - 208	207 - 208
41	14	2	ALA	ALA	LYS	LYS	1	B	C	207 - 208	207 - 208
42	14	2	ALA	ALA	LYS	LYS	1	C	E	207 - 208	207 - 208
43	15	2	LYS	LYS	LYS	LYS	3	A	A	209	209
44	15	2	LYS	LYS	LYS	LYS	3	B	C	209	209
45	15	2	LYS	LYS	LYS	LYS	3	C	E	209	209
46	16	2	GLY	GLY	LEU	LEU	1	A	A	210 - 213	210 - 213
47	16	2	GLY	GLY	LEU	LEU	1	B	C	210 - 213	210 - 213
48	16	2	GLY	GLY	LEU	LEU	1	C	E	210 - 213	210 - 213
49	17	2	TYR	TYR	GLU	GLU	3	A	A	214 - 215	214 - 215
50	17	2	TYR	TYR	GLU	GLU	3	B	C	214 - 215	214 - 215
51	17	2	TYR	TYR	GLU	GLU	3	C	E	214 - 215	214 - 215
52	18	2	LEU	LEU	GLY	GLY	1	A	A	216 - 220	216 - 220
53	18	2	LEU	LEU	GLY	GLY	1	B	C	216 - 220	216 - 220
54	18	2	LEU	LEU	GLY	GLY	1	C	E	216 - 220	216 - 220
55	19	2	PRO	PRO	PRO	PRO	2	A	A	221	221
56	19	2	PRO	PRO	PRO	PRO	2	B	C	221	221
57	19	2	PRO	PRO	PRO	PRO	2	C	E	221	221
58	20	2	VAL	VAL	PRO	PRO	1	A	A	222 - 228	222 - 228
59	20	2	VAL	VAL	PRO	PRO	1	B	C	222 - 228	222 - 228
60	20	2	VAL	VAL	PRO	PRO	1	C	E	222 - 228	222 - 228
61	21	2	LYS	LYS	LYS	LYS	3	A	A	229	229
62	21	2	LYS	LYS	LYS	LYS	3	B	C	229	229
63	21	2	LYS	LYS	LYS	LYS	3	C	E	229	229
64	22	2	VAL	VAL	ASN	ASN	1	A	A	230 - 233	230 - 233
65	22	2	VAL	VAL	ASN	ASN	1	B	C	230 - 233	230 - 233
66	22	2	VAL	VAL	ASN	ASN	1	C	E	230 - 233	230 - 233
67	23	2	GLN	GLN	GLN	GLN	2	A	A	234	234
68	23	2	GLN	GLN	GLN	GLN	2	B	C	234	234
69	23	2	GLN	GLN	GLN	GLN	2	C	E	234	234
70	24	2	VAL	VAL	TRP	TRP	1	A	A	235 - 254	235 - 254
71	24	2	VAL	VAL	TRP	TRP	1	B	C	235 - 254	235 - 254
72	24	2	VAL	VAL	TRP	TRP	1	C	E	235 - 254	235 - 254
73	25	2	ASP	ASP	ASP	ASP	4	A	A	255	255
74	25	2	ASP	ASP	ASP	ASP	4	B	C	255	255
75	25	2	ASP	ASP	ASP	ASP	4	C	E	255	255
76	26	2	THR	THR	THR	THR	1	A	A	256 - 258	256 - 258
77	26	2	THR	THR	THR	THR	1	B	C	256 - 258	256 - 258
78	26	2	THR	THR	THR	THR	1	C	E	256 - 258	256 - 258
79	27	2	PRO	PRO	PRO	PRO	2	A	A	259	259
80	27	2	PRO	PRO	PRO	PRO	2	B	C	259	259
81	27	2	PRO	PRO	PRO	PRO	2	C	E	259	259
82	28	2	PHE	PHE	TYR	TYR	1	A	A	260 - 261	260 - 261
83	28	2	PHE	PHE	TYR	TYR	1	B	C	260 - 261	260 - 261
84	28	2	PHE	PHE	TYR	TYR	1	C	E	260 - 261	260 - 261
85	29	2	SF4	SF4	F3S	F3S	1	A	G - H	265 - 266
86	29	2	SF4	SF4	F3S	F3S	1	B	P - Q	265 - 266
87	29	2	SF4	SF4	F3S	F3S	1	C	W - X	265 - 266
1	1	3	THR	THR	GLY	GLY	3	Q	B	7 - 14	7 - 14
2	1	3	THR	THR	GLY	GLY	3	R	D	7 - 14	7 - 14
3	1	3	THR	THR	GLY	GLY	3	S	F	7 - 14	7 - 14
4	2	3	PRO	PRO	VAL	VAL	1	Q	B	15 - 32	15 - 32
5	2	3	PRO	PRO	VAL	VAL	1	R	D	15 - 32	15 - 32
6	2	3	PRO	PRO	VAL	VAL	1	S	F	15 - 32	15 - 32
7	3	3	GLU	GLU	LYS	LYS	3	Q	B	33 - 40	33 - 40
8	3	3	GLU	GLU	LYS	LYS	3	R	D	33 - 40	33 - 40
9	3	3	GLU	GLU	LYS	LYS	3	S	F	33 - 40	33 - 40
10	4	3	ASP	ASP	LEU	LEU	1	Q	B	41 - 56	41 - 56
11	4	3	ASP	ASP	LEU	LEU	1	R	D	41 - 56	41 - 56
12	4	3	ASP	ASP	LEU	LEU	1	S	F	41 - 56	41 - 56
13	5	3	LYS	LYS	LYS	LYS	3	Q	B	57	57
14	5	3	LYS	LYS	LYS	LYS	3	R	D	57	57
15	5	3	LYS	LYS	LYS	LYS	3	S	F	57	57
16	6	3	GLY	GLY	PRO	PRO	1	Q	B	58 - 61	58 - 61
17	6	3	GLY	GLY	PRO	PRO	1	R	D	58 - 61	58 - 61
18	6	3	GLY	GLY	PRO	PRO	1	S	F	58 - 61	58 - 61
19	7	3	ARG	ARG	ARG	ARG	3	Q	B	62	62
20	7	3	ARG	ARG	ARG	ARG	3	R	D	62	62
21	7	3	ARG	ARG	ARG	ARG	3	S	F	62	62
22	8	3	GLY	GLY	VAL	VAL	1	Q	B	58 - 91	58 - 91
23	8	3	GLY	GLY	VAL	VAL	1	R	D	58 - 91	58 - 91
24	8	3	GLY	GLY	VAL	VAL	1	S	F	58 - 91	58 - 91
25	9	3	LYS	LYS	SER	SER	3	Q	B	92 - 94	92 - 94
26	9	3	LYS	LYS	SER	SER	3	R	D	92 - 94	92 - 94
27	9	3	LYS	LYS	SER	SER	3	S	F	92 - 94	92 - 94
28	10	3	ILE	ILE	HIS	HIS	1	Q	B	95 - 115	95 - 115
29	10	3	ILE	ILE	HIS	HIS	1	R	D	95 - 115	95 - 115
30	10	3	ILE	ILE	HIS	HIS	1	S	F	95 - 115	95 - 115
31	11	3	LEU	LEU	LEU	LEU	3	Q	B	116	116
32	11	3	LEU	LEU	LEU	LEU	3	R	D	116	116
33	11	3	LEU	LEU	LEU	LEU	3	S	F	116	116
34	12	3	VAL	VAL	VAL	VAL	1	Q	B	117 - 130	117 - 130
35	12	3	VAL	VAL	VAL	VAL	1	R	D	117 - 130	117 - 130
36	12	3	VAL	VAL	VAL	VAL	1	S	F	117 - 130	117 - 130
37	13	3	THR	THR	ALA	ALA	5	Q	B	131 - 133	131 - 133
38	13	3	THR	THR	ALA	ALA	5	R	D	131 - 133	131 - 133
39	13	3	THR	THR	ALA	ALA	5	S	F	131 - 133	131 - 133
40	14	3	ALA	ALA	ALA	ALA	4	Q	B	141 - 142	141 - 142
41	14	3	ALA	ALA	ALA	ALA	4	R	D	141 - 142	141 - 142
42	14	3	ALA	ALA	ALA	ALA	4	S	F	141 - 142	141 - 142
43	15	3	ALA	ALA	ALA	ALA	3	Q	B	151 - 159	151 - 159
44	15	3	ALA	ALA	ALA	ALA	3	R	D	151 - 159	151 - 159
45	15	3	ALA	ALA	ALA	ALA	3	S	F	151 - 159	151 - 159
46	16	3	LYS	LYS	LEU	LEU	3	Q	B	161 - 176	161 - 176
47	16	3	LYS	LYS	LEU	LEU	3	R	D	161 - 176	161 - 176
48	16	3	LYS	LYS	LEU	LEU	3	S	F	161 - 176	161 - 176
49	17	3	GLY	GLY	THR	THR	1	Q	B	177 - 180	177 - 180
50	17	3	GLY	GLY	THR	THR	1	R	D	177 - 180	177 - 180
51	17	3	GLY	GLY	THR	THR	1	S	F	177 - 180	177 - 180
52	18	3	ASN	ASN	LEU	LEU	3	Q	B	181 - 185	181 - 185
53	18	3	ASN	ASN	LEU	LEU	3	R	D	181 - 185	181 - 185
54	18	3	ASN	ASN	LEU	LEU	3	S	F	181 - 185	181 - 185
55	19	3	ALA	ALA	GLU	GLU	3	Q	B	190 - 196	190 - 196
56	19	3	ALA	ALA	GLU	GLU	3	R	D	190 - 196	190 - 196
57	19	3	ALA	ALA	GLU	GLU	3	S	F	190 - 196	190 - 196
58	20	3	VAL	VAL	TYR	TYR	1	Q	B	197 - 205	197 - 205
59	20	3	VAL	VAL	TYR	TYR	1	R	D	197 - 205	197 - 205
60	20	3	VAL	VAL	TYR	TYR	1	S	F	197 - 205	197 - 205
61	21	3	LEU	LEU	LYS	LYS	2	Q	B	206 - 214	206 - 214
62	21	3	LEU	LEU	LYS	LYS	2	R	D	206 - 214	206 - 214
63	21	3	LEU	LEU	LYS	LYS	2	S	F	206 - 214	206 - 214
64	22	3	ALA	ALA	TYR	TYR	1	Q	B	215 - 240	215 - 240
65	22	3	ALA	ALA	TYR	TYR	1	R	D	215 - 240	215 - 240
66	22	3	ALA	ALA	TYR	TYR	1	S	F	215 - 240	215 - 240
67	23	3	GLN	GLN	LYS	LYS	3	Q	B	241 - 278	241 - 278
68	23	3	GLN	GLN	LYS	LYS	3	R	D	241 - 278	241 - 278
69	23	3	GLN	GLN	LYS	LYS	3	S	F	241 - 278	241 - 278
70	24	3	ALA	ALA	THR	THR	2	Q	B	415 - 421	415 - 421
71	24	3	ALA	ALA	THR	THR	2	R	D	415 - 421	415 - 421
72	24	3	ALA	ALA	THR	THR	2	S	F	415 - 421	415 - 421
73	25	3	LEU	LEU	LEU	LEU	3	Q	B	422	422
74	25	3	LEU	LEU	LEU	LEU	3	R	D	422	422
75	25	3	LEU	LEU	LEU	LEU	3	S	F	422	422
76	26	3	GLY	GLY	MET	MET	1	Q	B	423 - 439	423 - 439
77	26	3	GLY	GLY	MET	MET	1	R	D	423 - 439	423 - 439
78	26	3	GLY	GLY	MET	MET	1	S	F	423 - 439	423 - 439
79	27	3	GLU	GLU	MET	MET	3	Q	B	440 - 446	440 - 446
80	27	3	GLU	GLU	MET	MET	3	R	D	440 - 446	440 - 446
81	27	3	GLU	GLU	MET	MET	3	S	F	440 - 446	440 - 446
82	28	3	LEU	LEU	LYS	LYS	3	Q	B	456 - 467	456 - 467
83	28	3	LEU	LEU	LYS	LYS	3	R	D	456 - 467	456 - 467
84	28	3	LEU	LEU	LYS	LYS	3	S	F	456 - 467	456 - 467
85	29	3	GLY	GLY	ILE	ILE	1	Q	B	468 - 483	468 - 483
86	29	3	GLY	GLY	ILE	ILE	1	R	D	468 - 483	468 - 483
87	29	3	GLY	GLY	ILE	ILE	1	S	F	468 - 483	468 - 483
88	30	3	ARG	ARG	ASN	ASN	3	Q	B	484 - 492	484 - 492
89	30	3	ARG	ARG	ASN	ASN	3	R	D	484 - 492	484 - 492
90	30	3	ARG	ARG	ASN	ASN	3	S	F	484 - 492	484 - 492
91	31	3	PHE	PHE	PRO	PRO	1	Q	B	493 - 505	493 - 505
92	31	3	PHE	PHE	PRO	PRO	1	R	D	493 - 505	493 - 505
93	31	3	PHE	PHE	PRO	PRO	1	S	F	493 - 505	493 - 505
94	32	3	ARG	ARG	ILE	ILE	3	Q	B	506 - 520	506 - 520
95	32	3	ARG	ARG	ILE	ILE	3	R	D	506 - 520	506 - 520
96	32	3	ARG	ARG	ILE	ILE	3	S	F	506 - 520	506 - 520
97	33	3	GLY	GLY	ALA	ALA	1	Q	B	521 - 525	521 - 525
98	33	3	GLY	GLY	ALA	ALA	1	R	D	521 - 525	521 - 525
99	33	3	GLY	GLY	ALA	ALA	1	S	F	521 - 525	521 - 525
100	34	3	PRO	PRO	ILE	ILE	1	Q	B	530 - 533	530 - 533
101	34	3	PRO	PRO	ILE	ILE	1	R	D	530 - 533	530 - 533
102	34	3	PRO	PRO	ILE	ILE	1	S	F	530 - 533	530 - 533
103	35	3	LEU	LEU	HIS	HIS	1	Q	B	534 - 549	534 - 549
104	35	3	LEU	LEU	HIS	HIS	1	R	D	534 - 549	534 - 549
105	35	3	LEU	LEU	HIS	HIS	1	S	F	534 - 549	534 - 549
106	36	3	FCO	FCO	MG	MG	1	Q	M - L	550 - 553
107	36	3	FCO	FCO	MG	MG	1	R	U - T	550 - 553
108	36	3	FCO	FCO	MG	MG	1	S	BA - AA	550 - 553
1	1	4	ASP	ASP	ASP	ASP	3	Q	B	279	279
2	1	4	ASP	ASP	ASP	ASP	3	R	D	279	279
3	1	4	ASP	ASP	ASP	ASP	3	S	F	279	279
4	2	4	TRP	TRP	THR	THR	1	Q	B	280 - 286	280 - 286
5	2	4	TRP	TRP	THR	THR	1	R	D	280 - 286	280 - 286
6	2	4	TRP	TRP	THR	THR	1	S	F	280 - 286	280 - 286
7	3	4	SER	SER	ASN	ASN	2	Q	B	287 - 288	287 - 288
8	3	4	SER	SER	ASN	ASN	2	R	D	287 - 288	287 - 288
9	3	4	SER	SER	ASN	ASN	2	S	F	287 - 288	287 - 288
10	4	4	TYR	TYR	ASP	ASP	1	Q	B	289 - 299	289 - 299
11	4	4	TYR	TYR	ASP	ASP	1	R	D	289 - 299	289 - 299
12	4	4	TYR	TYR	ASP	ASP	1	S	F	289 - 299	289 - 299
13	5	4	SER	SER	PRO	PRO	1	Q	B	301 - 302	301 - 302
14	5	4	SER	SER	PRO	PRO	1	R	D	301 - 302	301 - 302
15	5	4	SER	SER	PRO	PRO	1	S	F	301 - 302	301 - 302
16	6	4	LYS	LYS	GLY	GLY	1	Q	B	304 - 318	304 - 318
17	6	4	LYS	LYS	GLY	GLY	1	R	D	304 - 318	304 - 318
18	6	4	LYS	LYS	GLY	GLY	1	S	F	304 - 318	304 - 318
19	7	4	ARG	ARG	LEU	LEU	3	Q	B	319 - 329	319 - 329
20	7	4	ARG	ARG	LEU	LEU	3	R	D	319 - 329	319 - 329
21	7	4	ARG	ARG	LEU	LEU	3	S	F	319 - 329	319 - 329
22	8	4	ALA	ALA	LYS	LYS	3	Q	B	331 - 337	331 - 337
23	8	4	ALA	ALA	LYS	LYS	3	R	D	331 - 337	331 - 337
24	8	4	ALA	ALA	LYS	LYS	3	S	F	331 - 337	331 - 337
25	9	4	TYR	TYR	TYR	TYR	2	Q	B	338 - 341	338 - 341
26	9	4	TYR	TYR	TYR	TYR	2	R	D	338 - 341	338 - 341
27	9	4	TYR	TYR	TYR	TYR	2	S	F	338 - 341	338 - 341
28	10	4	GLY	GLY	GLY	GLY	4	Q	B	345	345
29	10	4	GLY	GLY	GLY	GLY	4	R	D	345	345
30	10	4	GLY	GLY	GLY	GLY	4	S	F	345	345
31	11	4	ASP	ASP	LYS	LYS	3	Q	B	346 - 348	346 - 348
32	11	4	ASP	ASP	LYS	LYS	3	R	D	346 - 348	346 - 348
33	11	4	ASP	ASP	LYS	LYS	3	S	F	346 - 348	346 - 348
34	12	4	HIS	HIS	TYR	TYR	2	Q	B	349 - 351	349 - 351
35	12	4	HIS	HIS	TYR	TYR	2	R	D	349 - 351	349 - 351
36	12	4	HIS	HIS	TYR	TYR	2	S	F	349 - 351	349 - 351
37	13	4	ASP	ASP	TYR	TYR	3	Q	B	352 - 359	352 - 359
38	13	4	ASP	ASP	TYR	TYR	3	R	D	352 - 359	352 - 359
39	13	4	ASP	ASP	TYR	TYR	3	S	F	352 - 359	352 - 359
40	14	4	LYS	LYS	HIS	HIS	3	Q	B	365 - 367	365 - 367
41	14	4	LYS	LYS	HIS	HIS	3	R	D	365 - 367	365 - 367
42	14	4	LYS	LYS	HIS	HIS	3	S	F	365 - 367	365 - 367
43	15	4	TYR	TYR	TYR	TYR	1	Q	B	368 - 376	368 - 376
44	15	4	TYR	TYR	TYR	TYR	1	R	D	368 - 376	368 - 376
45	15	4	TYR	TYR	TYR	TYR	1	S	F	368 - 376	368 - 376
46	16	4	ALA	ALA	ALA	ALA	1	Q	B	380 - 392	380 - 392
47	16	4	ALA	ALA	ALA	ALA	1	R	D	380 - 392	380 - 392
48	16	4	ALA	ALA	ALA	ALA	1	S	F	380 - 392	380 - 392
49	17	4	TYR	TYR	TYR	TYR	2	Q	B	393	393
50	17	4	TYR	TYR	TYR	TYR	2	R	D	393	393
51	17	4	TYR	TYR	TYR	TYR	2	S	F	393	393
52	18	4	ALA	ALA	LYS	LYS	3	Q	B	394 - 395	394 - 395
53	18	4	ALA	ALA	LYS	LYS	3	R	D	394 - 395	394 - 395
54	18	4	ALA	ALA	LYS	LYS	3	S	F	394 - 395	394 - 395
55	19	4	GLY	GLY	PHE	PHE	2	Q	B	396 - 400	396 - 400
56	19	4	GLY	GLY	PHE	PHE	2	R	D	396 - 400	396 - 400
57	19	4	GLY	GLY	PHE	PHE	2	S	F	396 - 400	396 - 400
58	20	4	LYS	LYS	LYS	LYS	3	Q	B	401	401
59	20	4	LYS	LYS	LYS	LYS	3	R	D	401	401
60	20	4	LYS	LYS	LYS	LYS	3	S	F	401	401
61	21	4	VAL	VAL	LEU	LEU	5	Q	B	407 - 408	407 - 408
62	21	4	VAL	VAL	LEU	LEU	5	R	D	407 - 408	407 - 408
63	21	4	VAL	VAL	LEU	LEU	5	S	F	407 - 408	407 - 408
64	22	4	LEU	LEU	PRO	PRO	3	Q	B	411 - 414	411 - 414
65	22	4	LEU	LEU	PRO	PRO	3	R	D	411 - 414	411 - 414
66	22	4	LEU	LEU	PRO	PRO	3	S	F	411 - 414	411 - 414

NCS ensembles :

ID
1
2
3
4

Components

Periplasmic [NiFe] hydrogenase ... , 2 types, 6 molecules A B C Q R S

#1: Protein

A B C Periplasmic [NiFe] hydrogenase small subunit / NiFe hydrogenlyase small chain

Details:

Mass: 28347.314 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Desulfovibrio fructosovorans (bacteria)
Strain: wild type / Gene: hydA / Production host: Desulfovibrio fructosovorans (bacteria) / References: UniProt: P18187, cytochrome-c3 hydrogenase

#2: Protein

Q R S Periplasmic [NiFe] hydrogenase large subunit / NiFe hydrogenlyase large chain

Details:

Mass: 59836.375 Da / Num. of mol.: 3 / Mutation: V74I, L122F
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Desulfovibrio fructosovorans (bacteria)
Strain: wild type / Gene: hydB / Production host: Desulfovibrio fructosovorans (bacteria) / References: UniProt: P18188, cytochrome-c3 hydrogenase

Non-polymers , 7 types, 1036 molecules

#3: Chemical

A-265 A-267 B-265 B-267 C-265 C-267
ChemComp-SF4 / IRON/SULFUR CLUSTER

Details:

Mass: 351.640 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: Fe₄S₄

#4: Chemical

A-266 B-266 C-266 ChemComp-F3S / FE3-S4 CLUSTER

Details:

Mass: 295.795 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: Fe₃S₄

#5: Chemical

A-271 Q-561 Q-562 R-563 S-561
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL

Details:

Mass: 92.094 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: C₃H₈O₃

#6: Chemical

Q-551 R-551 S-551 ChemComp-NI / NICKEL (II) ION

Details:

Mass: 58.693 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: Ni

#7: Chemical

Q-553 R-553 S-553 ChemComp-MG / MAGNESIUM ION

Details:

Mass: 24.305 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: Mg

#8: Chemical

Q-550 R-550 S-550 ChemComp-FCO / CARBONMONOXIDE-(DICYANO) IRON

Details:

Mass: 135.890 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C₃FeN₂O

#9: Water

ChemComp-HOH / water

Details:

Mass: 18.015 Da / Num. of mol.: 1013 / Source method: isolated from a natural source / Formula: H₂O

Details

• Has protein modification: Y

Experimental details

Experiment

• Experiment: Method: X-RAY DIFFRACTION / Number of used crystals: 1

Sample preparation

• Crystal: Density Matthews: 2.09 ų/Da / Density % sol: 41.2 %
• Crystal grow: Temperature: 293 K / Method: vapor diffusion, hanging drop / pH: 6 ; Details: PEG6000, GLYCEROL, PH6.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K

Data collection

• Diffraction: Mean temperature: 100 K
• Diffraction source: Source: SYNCHROTRON / Site: ESRF / Beamline: ID14-4 / Wavelength: 1 / Wavelength: 0.934 Å
• Detector: Type: ADSC QUANTUM 315r / Detector: CCD / Date: Jan 26, 2008 / Details: MIRRORS
• Radiation: Monochromator: SI(111) MONOCHROMATOR / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray

Radiation wavelength

ID	Wavelength (Å)	Relative weight
1	1	1
2	0.934	1

• Reflection: Resolution: 2.18→20 Å / Num. all: 115860 / Num. obs: 108157 / % possible obs: 93.3 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 2.6 % / B_iso Wilson estimate: 27 Ų / R_merge(I) obs: 0.07 / R_sym value: 0.07 / Net I/σ(I): 7.2
• Reflection shell: Resolution: 2.18→2.3 Å / Redundancy: 2.2 % / R_merge(I) obs: 0.198 / Mean I/σ(I) obs: 3.3 / Num. unique all: 17163 / R_sym value: 0.198 / % possible all: 71

Processing

Software

Name	Version	Classification
AMoRE		phasing
REFMAC	5.2.0019	refinement
ADSC	Quantum	data collection
XDS		data reduction
XDS		data scaling

Refinement

Method to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 1YQW

1yqw

Resolution: 2.2→20 Å / Cor.coef. F_o:F_c: 0.932 / Cor.coef. F_o:F_c free: 0.906 / SU B: 11.41 / SU ML: 0.15 / Cross valid method: THROUGHOUT / σ(F): 0 / σ(I): 0 / ESU R: 0.387 / ESU R Free: 0.223 / Stereochemistry target values: MAXIMUM LIKELIHOOD

	Rfactor	Num. reflection	% reflection	Selection details
Rfree	0.22223	5404	5 %	RANDOM
Rwork	0.18493	-	-	-
obs	0.18677	101754	94.24 %	-
all	-	0	-	-

• Solvent computation: Ion probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: BABINET MODEL WITH MASK

Displacement parameters

B_iso mean: 15.032 Ų

	Baniso -1	Baniso -2	Baniso -3
1-	3.07 Å2	0 Å2	0.21 Å2
2-	-	-0.93 Å2	0 Å2
3-	-	-	-2.13 Å2

Refinement step

Cycle: LAST / Resolution: 2.2→20 Å

	Protein	Nucleic acid	Ligand	Solvent	Total
Num. atoms	18418	0	126	1013	19557

Refine LS restraints

Refine-ID	Type	Dev ideal	Dev ideal target	Number
X-RAY DIFFRACTION	r_bond_refined_d	0.012	0.022	19146
X-RAY DIFFRACTION	r_bond_other_d
X-RAY DIFFRACTION	r_angle_refined_deg	1.266	1.965	25958
X-RAY DIFFRACTION	r_angle_other_deg
X-RAY DIFFRACTION	r_dihedral_angle_1_deg	9.521	5.025	2417
X-RAY DIFFRACTION	r_dihedral_angle_2_deg	34.162	24.413	775
X-RAY DIFFRACTION	r_dihedral_angle_3_deg	14.12	15	3040
X-RAY DIFFRACTION	r_dihedral_angle_4_deg	14.543	15	72
X-RAY DIFFRACTION	r_chiral_restr	0.085	0.2	2826
X-RAY DIFFRACTION	r_gen_planes_refined	0.004	0.02	14457
X-RAY DIFFRACTION	r_gen_planes_other
X-RAY DIFFRACTION	r_nbd_refined	0.196	0.2	9498
X-RAY DIFFRACTION	r_nbd_other
X-RAY DIFFRACTION	r_nbtor_refined	0.3	0.2	12921
X-RAY DIFFRACTION	r_nbtor_other
X-RAY DIFFRACTION	r_xyhbond_nbd_refined	0.147	0.2	1358
X-RAY DIFFRACTION	r_xyhbond_nbd_other
X-RAY DIFFRACTION	r_metal_ion_refined
X-RAY DIFFRACTION	r_metal_ion_other
X-RAY DIFFRACTION	r_symmetry_vdw_refined	0.215	0.2	97
X-RAY DIFFRACTION	r_symmetry_vdw_other
X-RAY DIFFRACTION	r_symmetry_hbond_refined	0.21	0.2	35
X-RAY DIFFRACTION	r_symmetry_hbond_other
X-RAY DIFFRACTION	r_symmetry_metal_ion_refined
X-RAY DIFFRACTION	r_symmetry_metal_ion_other
X-RAY DIFFRACTION	r_mcbond_it	0.401	1.5	12431
X-RAY DIFFRACTION	r_mcbond_other
X-RAY DIFFRACTION	r_mcangle_it	0.602	3	19307
X-RAY DIFFRACTION	r_scbond_it	0.72	2	7778
X-RAY DIFFRACTION	r_scangle_it	1.094	4.5	6570
X-RAY DIFFRACTION	r_rigid_bond_restr
X-RAY DIFFRACTION	r_sphericity_free
X-RAY DIFFRACTION	r_sphericity_bonded

Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION

Ens-ID	Dom-ID	Auth asym-ID	Number	Type	Rms dev position (Å)	Weight position
1	1	A	951	tight positional	0.05	0.06
1	2	B	951	tight positional	0.04	0.06
1	3	C	951	tight positional	0.05	0.06
2	1	A	603	tight positional	0.04	0.06
2	2	B	603	tight positional	0.04	0.06
2	3	C	603	tight positional	0.05	0.06
3	1	Q	2297	tight positional	0.05	0.06
3	2	R	2297	tight positional	0.05	0.06
3	3	S	2297	tight positional	0.05	0.06
4	1	Q	653	tight positional	0.05	0.06
4	2	R	653	tight positional	0.05	0.06
4	3	S	653	tight positional	0.05	0.06
1	1	A	46	medium positional	0.26	0.2
1	2	B	46	medium positional	0.25	0.2
1	3	C	46	medium positional	0.47	0.2
2	1	A	36	medium positional	0.11	0.2
2	2	B	36	medium positional	0.1	0.2
2	3	C	36	medium positional	0.1	0.2
3	1	Q	79	medium positional	0.14	0.2
3	2	R	79	medium positional	0.12	0.2
3	3	S	79	medium positional	0.13	0.2
4	1	Q	90	medium positional	0.11	0.2
4	2	R	90	medium positional	0.1	0.2
4	3	S	90	medium positional	0.12	0.2
1	1	A	126	loose positional	0.41	5
1	2	B	126	loose positional	0.57	5
1	3	C	126	loose positional	0.42	5
2	1	A	88	loose positional	0.29	5
2	2	B	88	loose positional	0.39	5
2	3	C	88	loose positional	0.37	5
3	1	Q	532	loose positional	0.44	5
3	2	R	532	loose positional	0.44	5
3	3	S	532	loose positional	0.52	5
4	1	Q	160	loose positional	0.43	5
4	2	R	160	loose positional	0.35	5
4	3	S	160	loose positional	0.43	5
1	1	A	951	tight thermal	0.29	1
1	2	B	951	tight thermal	0.27	1
1	3	C	951	tight thermal	0.28	1
2	1	A	603	tight thermal	0.32	1
2	2	B	603	tight thermal	0.31	1
2	3	C	603	tight thermal	0.27	1
3	1	Q	2297	tight thermal	0.3	1
3	2	R	2297	tight thermal	0.28	1
3	3	S	2297	tight thermal	0.27	1
4	1	Q	653	tight thermal	0.31	1
4	2	R	653	tight thermal	0.25	1
4	3	S	653	tight thermal	0.27	1
1	1	A	46	medium thermal	0.58	2
1	2	B	46	medium thermal	0.48	2
1	3	C	46	medium thermal	0.55	2
2	1	A	36	medium thermal	0.59	2
2	2	B	36	medium thermal	0.6	2
2	3	C	36	medium thermal	0.5	2
3	1	Q	79	medium thermal	0.76	2
3	2	R	79	medium thermal	0.53	2
3	3	S	79	medium thermal	0.78	2
4	1	Q	90	medium thermal	0.66	2
4	2	R	90	medium thermal	0.61	2
4	3	S	90	medium thermal	0.48	2
1	1	A	126	loose thermal	1.51	10
1	2	B	126	loose thermal	1.55	10
1	3	C	126	loose thermal	1.83	10
2	1	A	88	loose thermal	1.3	10
2	2	B	88	loose thermal	1.42	10
2	3	C	88	loose thermal	1	10
3	1	Q	532	loose thermal	1.16	10
3	2	R	532	loose thermal	1.08	10
3	3	S	532	loose thermal	1.03	10
4	1	Q	160	loose thermal	1.01	10
4	2	R	160	loose thermal	1.04	10
4	3	S	160	loose thermal	1.02	10

LS refinement shell

Resolution: 2.2→2.256 Å / Total num. of bins used: 20

	Rfactor	Num. reflection	% reflection
Rfree	0.279	356	-
Rwork	0.222	6639	-
obs	-	-	83.9 %