Li, Huilin, Wells, Stephen A., Jimenez-Roldan, J. Emilio, Römer, Rudolf A., Zhao, Yao, (Researcher in chemistry), Sadler, P. J. and O'Connor, Peter B. (2012) Protein flexibility is key to cisplatin crosslinking in calmodulin. Protein Science, Volume 21 (Number 9). pp. 1269-1279. doi:10.1002/pro.2111

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Abstract

Chemical crosslinking in combination with Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) has significant potential for studying protein structures and proteinprotein interactions. Previously, cisplatin has been shown to be a crosslinker and crosslinks multiple methionine (Met) residues in apo-calmodulin (apo-CaM). However, the inter-residue distances obtained from nuclear magnetic resonance structures are inconsistent with the measured distance constraints by crosslinking. Met residues lie too far apart to be crosslinked by cisplatin. Here, by combining FTICR MS with a novel computational flexibility analysis, the flexible nature of the CaM structure is found to be key to cisplatin crosslinking in CaM. It is found that the side chains of Met residues can be brought together by flexible motions in both apo-CaM and calcium-bound CaM (Ca4-CaM). The possibility of cisplatin crosslinking Ca4-CaM is then confirmed by MS data. Therefore, flexibility analysis as a fast and low-cost computational method can be a useful tool for predicting crosslinking pairs in protein crosslinking analysis and facilitating MS data analysis. Finally, flexibility analysis also indicates that the crosslinking of platinum to pairs of Met residues will effectively close the nonpolar groove and thus will likely interfere with the binding of CaM to its protein targets, as was proved by comparing assays for cisplatin-modified/unmodified CaM binding to melittin. Collectively, these results suggest that cisplatin crosslinking of apo-CaM or Ca4-CaM can inhibit the ability of CaM to recognize its target proteins, which may have important implications for understanding the mechanism of tumor resistance to platinum anticancer drugs.

Item Type:	Journal Article
Subjects:	Q Science > QD Chemistry Q Science > QH Natural history > QH301 Biology
Divisions:	Faculty of Science > Chemistry Faculty of Science > Centre for Complexity Science Faculty of Science > Physics
Library of Congress Subject Headings (LCSH):	Proteins -- Crosslinking, Cisplatin, Fourier transform nuclear magnetic resonance spectroscopy, Protein-protein interactions
Journal or Publication Title:	Protein Science
Publisher:	John Wiley & Sons Ltd.
ISSN:	0961-8368
Official Date:	September 2012
Dates:	Date Event September 2012 Published
Date of first compliant deposit:	23 December 2015
Volume:	Volume 21
Number:	Number 9
Page Range:	pp. 1269-1279
DOI:	10.1002/pro.2111
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	University of Warwick Postgraduate Research Scholarship, University of Warwick. Dept. of Chemistry, Leverhulme Trust (LT), National Institutes of Health (U.S.) (NIH), European Research Council (ERC), Engineering and Physical Sciences Research Council (EPSRC), University of Warwick. Protein Biology and Biophysics Network
Grant number:	R01 GM078293 (NIH/NIGMS) ; 247450 (ERC) ; EP/F034210/1, BP/G006792 (EPSRC)

Data sourced from Thomson Reuters' Web of Knowledge

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