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BMRB: 19891

2MNG

A Mechanism for the cAMP-Dependent Auto-Inhibition of HCN

Authors

Akimoto, M., Zhang, Z., Boulton, S., Selvaratnam, R., VanSchouwen, B., Gloyd, M., Accili, E.A., Lange, O.F., Melacini, G.

Assembly

Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4

Entity

1. Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4 (polymer, Thiol state: all free), 131 monomers, 14967.08 Da Detail

NSEEIINFNC RKLVASMPLF ANADPNFVTS MLTKLRFEVF QPGDYIIREG TIGKKMYFIQ HGVVSVLTKG NKETKLADGS YFGEICLLTR GRRTASVRAD TYCRLYSLSV DNFNEVLEEY PMMRRAFETV A

Formula weight

14967.08 Da

Source organism

Homo sapiens

Exptl. method

solution NMR

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 96.9 %, Completeness: 77.7 %, Completeness (bb): 93.2 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	77.7 % (1206 of 1553)	75.4 % (608 of 806)	78.2 % (477 of 610)	88.3 % (121 of 137)
Backbone	93.2 % (725 of 778)	93.6 % (249 of 266)	92.2 % (355 of 385)	95.3 % (121 of 127)
Sidechain	66.4 % (596 of 898)	66.5 % (359 of 540)	68.1 % (237 of 348)	0.0 % (0 of 10)
Aromatic	11.3 % (16 of 142)	12.7 % (9 of 71)	9.9 % (7 of 71)
Methyl	74.7 % (109 of 146)	67.1 % (49 of 73)	82.2 % (60 of 73)

1. Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4

NSEEIINFNC RKLVASMPLF ANADPNFVTS MLTKLRFEVF QPGDYIIREG TIGKKMYFIQ HGVVSVLTKG NKETKLADGS YFGEICLLTR GRRTASVRAD TYCRLYSLSV DNFNEVLEEY PMMRRAFETV A

Show sample condition

Sample #1

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 300 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	DTT	natural abundance	1 mM
2	MES	natural abundance	20 mM
3	potassium chloride	natural abundance	100 mM
4	EDTA	natural abundance	2 mM
5	EGTA	natural abundance	2 mM
6	sodium azide	natural abundance	0.02 %
7	Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4	[U-13C; U-15N]	0.5 mM
8	D2O	natural abundance	95 %
9	H2O	natural abundance	5 %

Sample #2

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 300 K, pH 6.5

#	Name	Isotope labeling	Concentration
10	DTT	natural abundance	1 mM
11	MES	natural abundance	20 mM
12	potassium chloride	natural abundance	100 mM
13	EDTA	natural abundance	1 mM
14	sodium azide	natural abundance	0.02 %
15	Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4	[U-15N]	0.2 mM
16	Pf1 phage	natural abundance	6 mg/mL
17	D2O	natural abundance	95 %
18	H2O	natural abundance	5 %

Sample #3

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 300 K, pH 6.5

#	Name	Isotope labeling	Concentration
19	DTT	natural abundance	1 mM
20	MES	natural abundance	20 mM
21	potassium chloride	natural abundance	100 mM
22	EDTA	natural abundance	1 mM
23	sodium azide	natural abundance	0.02 %
24	Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4	[U-15N]	0.2 mM
25	D2O	natural abundance	95 %
26	H2O	natural abundance	5 %

LACS Plot; CA

Atom type	Mol. common name	Atom group	Value	Ref. method	Ref. type	Shift ratio
¹H	water	protons	4.75 ppm	internal	direct	1.0

Referencing offset: -0.17 ppm, Outliers: 2 Detail

LACS Plot; CB

Atom type	Mol. common name	Atom group	Value	Ref. method	Ref. type	Shift ratio
¹H	water	protons	4.75 ppm	internal	direct	1.0

Referencing offset: -0.17 ppm, Outliers: 2 Detail

LACS Plot; HA

Atom type	Mol. common name	Atom group	Value	Ref. method	Ref. type	Shift ratio
¹H	water	protons	4.75 ppm	internal	direct	1.0

Referencing offset: -0.04 ppm, Outliers: 1 Detail

LACS Plot; CO

Atom type	Mol. common name	Atom group	Value	Ref. method	Ref. type	Shift ratio
¹H	water	protons	4.75 ppm	internal	direct	1.0

Referencing offset: 0.22 ppm, Outliers: 3 Detail

Protein Blocks Logo

Calculated from 10 models in PDB: 2MNG, Strand ID: A Detail

Release date

2016-06-16

Citation

A mechanism for the auto-inhibition of hyperpolarization-activated cyclic nucleotide-gated (HCN) channel opening and its relief by cAMP
Akimoto, M., Zhang, Z., Boulton, S., Selvaratnam, R., VanSchouwen, B., Gloyd, M., Accili, E.A., Lange, O.F., Melacini, G.
J. Biol. Chem. (2014), 289, 22205-22220, PubMed 24878962 , DOI 10.1074/jbc.M114.572164 , Abstract

Related entities 1. Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4, : 1 : 15 : 260 entities Detail

Interaction partners 1. Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4, : 2 interactors Detail

More details for 2 interactors identified by 2 citations

Experiments performed 10 experiments Detail

1 2D 1H-15N HSQC