Isotope-edited Fourier transform infrared spectroscopy studies of calmodulin's interaction with its target peptides

Biochemistry
M ZhangH J Vogel

Abstract

The ubiquitous calcium-binding protein calmodulin (CaM) regulates a wide variety of cellular events by binding to and activating many distinct target enzymes. The CaM-binding domains of most of these enzymes are contained in a contiguous stretch of amino acids with a length of approximately 20 residues. In this work, we have used "isotope-edited" Fourier transform infrared spectroscopy to study the interaction of CaM with synthetic peptides resembling the CaM-binding domains of myosin light chain kinase (MLCK), constitutive nitric oxide synthase (cNOS), and caldesmon (CaD). Uniform labeling of CaM with carbon-13 causes the amide I band of the protein to shift approximately 55 cm-1 to lower frequency in D2O, leaving a clear window in the infrared spectrum for observing the amide I band of the unlabeled target peptides. Upon complex formation, the amide I bands of the CaM-binding domains of MLCK and cNOS shift 4 cm-1 toward higher frequency (to approximately 1648 cm-1), and have a narrower bandwidth compared to the peptide in aqueous solution. These spectral changes and the fact that the infrared spectra of these two peptides in their complex with CaM closely resemble those recorded in a mixture of D2O and the helix inducing solv...Continue Reading

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Citations

Aug 1, 1995·Molecular and Cellular Biochemistry·H J Vogel, M Zhang
May 24, 2006·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·R Sean PeacockHans J Vogel
Sep 6, 2013·Analytical Chemistry·Ying WangJian-Ding Qiu
May 18, 2013·Chemical Reviews·Heejae Kim, Minhaeng Cho
Dec 9, 2010·Journal of the American Chemical Society·Gai LiuNoel D Lazo
Apr 8, 2006·The Journal of Physical Chemistry. B·Jianping WangRobin M Hochstrasser
Aug 23, 2011·Journal of Structural Biology·Ganesh Shanmugam, Prasad L Polavarapu
Feb 19, 2011·Biochemical and Biophysical Research Communications·Marianne T BauerSarah A Petty
Jan 8, 2008·Journal of Molecular Biology·Heinz FabianDieter Naumann
Oct 13, 2006·Journal of Peptide Science : an Official Publication of the European Peptide Society·Valery V AndrushchenkoElmar J Prenner
Oct 13, 2011·Proteins·Liliana Del Valle SosaBelinda Pastrana-Ríos
Aug 19, 2007·Biochimica Et Biophysica Acta·Andreas Barth
Jun 2, 2005·Biochimica Et Biophysica Acta·Jane M VanderkooiBogumil Zelent
Jul 22, 2014·Journal of Pharmaceutical and Biomedical Analysis·Matthias D'HondtBart De Spiegeleer
Dec 4, 2012·Biochimica Et Biophysica Acta·Masayuki NaraMasaru Tanokura
Jan 10, 2008·Biochemical and Biophysical Research Communications·Masayuki Nara, Masaru Tanokura
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Mar 24, 2018·Chemical Science·Maurizio BaldassarreAndreas Barth
Jul 21, 2006·The Journal of Physical Chemistry. B·Chong Fang, Robin M Hochstrasser

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