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Title: Structural and Energetic Analysis of Activiation by a Cyclic Nucleotide Binding Domain

Abstract

MlotiK1 is a prokaryotic homolog of cyclic-nucleotide-dependent ion channels that contains an intracellular C-terminal cyclic nucleotide binding (CNB) domain. X-ray structures of the CNB domain have been solved in the absence of ligand and bound to cAMP. Both the full-length channel and CNB domain fragment are easily expressed and purified, making MlotiK1 a useful model system for dissecting activation by ligand binding. We have used X-ray crystallography to determine three new MlotiK1 CNB domain structures: a second apo configuration, a cGMP-bound structure, and a second cAMP-bound structure. In combination, the five MlotiK1 CNB domain structures provide a unique opportunity for analyzing, within a single protein, the structural differences between the apo state and the bound state, and the structural variability within each state. With this analysis as a guide, we have probed the nucleotide selectivity and importance of specific residue side chains in ligand binding and channel activation. These data help to identify ligand-protein interactions that are important for ligand dependence in MlotiK1 and, more globally, in the class of nucleotide-dependent proteins.

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
959733
Report Number(s):
BNL-82719-2009-JA
Journal ID: ISSN 0022-2836; JMOBAK; TRN: US201016%%877
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
Journal of Molecular Biology
Additional Journal Information:
Journal Volume: 381; Journal Issue: 3; Journal ID: ISSN 0022-2836
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; BOUND STATE; CHAINS; CONFIGURATION; CRYSTALLOGRAPHY; DOMAIN STRUCTURE; NUCLEOTIDES; PROTEINS; RESIDUES; national synchrotron light source

Citation Formats

Altieri, S, Clayton, G, Silverman, W, Olivares, A, De La Cruz, E, Thomas, L, and Morais-Cabral, J. Structural and Energetic Analysis of Activiation by a Cyclic Nucleotide Binding Domain. United States: N. p., 2008. Web. doi:10.1016/j.jmb.2008.06.011.
Altieri, S, Clayton, G, Silverman, W, Olivares, A, De La Cruz, E, Thomas, L, & Morais-Cabral, J. Structural and Energetic Analysis of Activiation by a Cyclic Nucleotide Binding Domain. United States. doi:10.1016/j.jmb.2008.06.011.
Altieri, S, Clayton, G, Silverman, W, Olivares, A, De La Cruz, E, Thomas, L, and Morais-Cabral, J. Tue . "Structural and Energetic Analysis of Activiation by a Cyclic Nucleotide Binding Domain". United States. doi:10.1016/j.jmb.2008.06.011.
@article{osti_959733,
title = {Structural and Energetic Analysis of Activiation by a Cyclic Nucleotide Binding Domain},
author = {Altieri, S and Clayton, G and Silverman, W and Olivares, A and De La Cruz, E and Thomas, L and Morais-Cabral, J},
abstractNote = {MlotiK1 is a prokaryotic homolog of cyclic-nucleotide-dependent ion channels that contains an intracellular C-terminal cyclic nucleotide binding (CNB) domain. X-ray structures of the CNB domain have been solved in the absence of ligand and bound to cAMP. Both the full-length channel and CNB domain fragment are easily expressed and purified, making MlotiK1 a useful model system for dissecting activation by ligand binding. We have used X-ray crystallography to determine three new MlotiK1 CNB domain structures: a second apo configuration, a cGMP-bound structure, and a second cAMP-bound structure. In combination, the five MlotiK1 CNB domain structures provide a unique opportunity for analyzing, within a single protein, the structural differences between the apo state and the bound state, and the structural variability within each state. With this analysis as a guide, we have probed the nucleotide selectivity and importance of specific residue side chains in ligand binding and channel activation. These data help to identify ligand-protein interactions that are important for ligand dependence in MlotiK1 and, more globally, in the class of nucleotide-dependent proteins.},
doi = {10.1016/j.jmb.2008.06.011},
journal = {Journal of Molecular Biology},
issn = {0022-2836},
number = 3,
volume = 381,
place = {United States},
year = {2008},
month = {1}
}