Interaction of calmodulin with its binding domain of rat cerebellar nitric oxide synthase. A multinuclear NMR study.

The Journal of Biological Chemistry
M ZhangH J Vogel

Abstract

The intercellular messenger nitric oxide is produced through the action of nitric oxide synthases, a class of enzymes that is regulated by calcium-calmodulin (CaM). In this work, the interaction of CaM with a 23-amino-acid residue synthetic peptide, encompassing the CaM-binding domain of constitutive rat cerebellar nitric oxide synthase (cNOS), was investigated by various NMR methods. Cadmium-113 NMR studies showed that binding of the cNOS peptide increased the affinity of CaM for metal ions and induced interdomain cooperativity in metal ion binding as earlier observed for complexes of CaM with myosin light chain kinase (MLCK) peptides. By using specific isotopically labeled [13C]methyl-Met and selenomethionine-substituted CaM in two-dimensional proton-detected 13C and 77Se NMR studies, we obtained evidence for the involvement of the Met residues of CaM in the binding of the cNOS peptide. These residues form two hydrophobic surface areas on CaM, and they are also involved in the binding of other target proteins. A nitroxide spin-labeled version of the cNOS peptide caused broadening only for NMR resonances in the N-terminal half of CaM, showing that the peptide binds with a C to N orientation to the N- and C-terminal domains of ...Continue Reading

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Citations

Aug 1, 1995·Molecular and Cellular Biochemistry·H J Vogel, M Zhang
Jun 8, 2001·Biochemical and Biophysical Research Communications·S J LeeJ T Stull
Oct 28, 2011·The Journal of Chemical Physics·Ali Rana AtilganCanan Atilgan
Sep 25, 2007·Biochimica Et Biophysica Acta·Donald E SprattJ Guy Guillemette
Mar 30, 2001·The Journal of Biological Chemistry·R D BrokxG I Makhatadze
May 13, 2004·The Journal of Biological Chemistry·Elena NewmanJ Guy Guillemette
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Sep 4, 2021·Frontiers in Molecular Biosciences·Kari GaalswykJustin L MacCallum
Aug 31, 2002·Biophysical Journal·Andriyka L PapishHans J Vogel

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