NMR studies of the methionine methyl groups in calmodulin

FEBS Letters

Siivari, Kirsi¹  Palmer, Arthur G.²  Zhang, Mingjie¹  Vogel, Hans J.¹ 

[1] Department of Biological Sciences, University of Calgary, 2500 University Drive N.W., Calgary, Alberta, T2N 1N4, Canada;Department of Biochemistry and Molecular Biophysics, Columbia University, 630 W. 168th St., New York, NY 10032, USA

关键词: Calmodulin;    Methionine;    Calcium;    Mutant;    Dynamics;    CaM;    calmodulin;    MLCK;    myosin light chain kinase;    NMR;    nuclear magnetic resonance;    HMQC;    heteronuclear multiple quantum coherence;    nOe;    nuclear Overhauser effect;    2D;    two-dimensional;    wt-CaM;    wild type calmodulin;   

DOI  :  10.1016/0014-5793(95)00504-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Calmodulin (CaM) is a ubiquitous Ca²⁺-binding protein that can regulate a wide variety of cellular events. The protein contains 9 Met out of a total of 148 amino acid residues. The binding of Ca²⁺ to CaM induces conformational changes and exposes two Met-rich hydrophobic surfaces which provide the main protein-protein contact areas when CaM interacts with its target enzymes. Two-dimensional (¹H,¹³C)-heteronuclear multiple quantum coherence (HMQC) NMR spectroscopy was used to study selectively ¹³C-isotope labelled Met methyl groups in apo-CaM, Ca²⁺-CaM and a complex of CaM with the CaM-binding domain of skeletal muscle Myosin Light Chain Kinase (MLCK). The resonance assignment of the Met methyl groups in these three functionally different states were obtained by site-directed mutagenesis (Met→Leu). Chemical shift changes indicate that the methyl groups of the Met residues are in different environments in apo-, calcium-, and MLCK-bound-CaM. The T ₁ relaxation rates of the individual Met methyl carbons in the three forms of CaM indicate that those in Ca²⁺-CaM have the highest mobility. Our results also suggest that the methyl groups of the unbranched Met sidechains in general are more flexible than those of aliphatic amino acid residues such as Leu and Ile.

【 授权许可】

Unknown