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Molecular and Cellular Biochemistry

, Volume 149, Issue 1, pp 3–15 | Cite as

Protein engineering and NMR studies of calmodulin

  • Hans J. Vogel
  • Mingjie Zhang
Article

Abstract

The calcium regulatory protein calmodulin (CaM) plays a role as an on-off switch in the activation of many enzymes and proteins. CaM has a dumbbell shaped structure with two folded domains, which are connected by a flexible linker in solution. The calmodulin-binding domains of the target proteins are contained in 20 residue long amino acid sequences, that share no obvious amino acid sequence homology. In this contribution, we discuss the features of CaM, which allow it to be rather promiscous, and bind effectively to all these distinct domains. In particular, we describe the role of the methionine-rich hydrophobic surfaces of the protein in providing a malleable and sticky surface for binding many hydrophobic peptides. The enzyme activation properties of various Met→Leu mutants of CaM are discussed. In addition, the role of the flexible linker region that connects the two domains is also analyzed. Finally, we describe various NMR and spectroscopic experiments that aid in determining the CaM-bound structures of synthetic peptides containing various CaM-binding domains. All structures analyzed to date are α-helical when bound to CaM, and they interact with CaM only through amino acid sidechains. This form of protein-protein interaction is rather unique, and may contribute to CaM's capacity to bind effectively to such a wide range of distinct partners.

Key words

calmodulin calcium NMR studies methionine protein-protein interaction 

Abbreviations

CaM

calmodulin

CD

circular dichroism

cNOS

constitutive Nitric Oxide Synthase

FTIR

Fourier Transform Infrared Spectroscopy

iNOS

inducible Nitric Oxide Synthase

MLCK

Myosin Light Chain Kinase

NMR

Nuclear Magnetic Resonance

nOe

nuclear Overhauser effect

NOESY

two dimensional Nuclear Overhauser Effect Spectroscopy

PDE

cyclic nucleotide phosphodiesterase

SeMet

selenomethionine

TFE

trifluoroethanol

TOCSY

two dimensional Total Correlation Spectroscopy

trnOe

transferred nuclear Overhauser effect

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Hans J. Vogel
    • 1
  • Mingjie Zhang
    • 1
  1. 1.Department of Biological SciencesThe University of CalgaryCalgaryCanada

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