Abstract
The enzyme nitric oxide synthase (NOS) is exquisitely regulated in vivo by the Ca2+ sensor protein calmodulin (CaM) to control production of NO, a key signaling molecule and cytotoxin. The differential activation of NOS isozymes by CaM has remained enigmatic, despite extensive research. Here, the crystallographic structure of Ca2+-loaded CaM bound to a 20 residue peptide comprising the endothelial NOS (eNOS) CaM-binding region establishes their individual conformations and intermolecular interactions, and suggests the basis for isozyme-specific differences. The α-helical eNOS peptide binds in an antiparallel orientation to CaM through extensive hydrophobic interactions. Unique NOS interactions occur with: (i) the CaM flexible central linker, explaining its importance in NOS activation; and (ii) the CaM C-terminus, explaining the NOS-specific requirement for a bulky, hydrophobic residue at position 144. This binding mode expands mechanisms for CaM-mediated activation, explains eNOS deactivation by Thr495 phosphorylation, and implicates specific hydrophobic residues in the Ca2+ independence of inducible NOS.
Original language | English (US) |
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Pages (from-to) | 766-775 |
Number of pages | 10 |
Journal | EMBO Journal |
Volume | 22 |
Issue number | 4 |
DOIs | |
State | Published - Feb 17 2003 |
Externally published | Yes |
Keywords
- Calcium
- Calmodulin
- Crystal structure
- Intermolecular interaction
- Nitric oxide synthase
ASJC Scopus subject areas
- General Neuroscience
- Molecular Biology
- General Biochemistry, Genetics and Molecular Biology
- General Immunology and Microbiology