Local dynamics of the M13 major coat protein in different membrane-mimicking systems

Biochemistry
D StoparM A Hemminga

Abstract

The local environment of the transmembrane and C-terminal domain of M13 major coat protein was probed by site-directed ESR spin labeling when the protein was introduced into three membrane-mimicking systems, DOPC vesicles, sodium cholate micelles, and SDS micelles. For this purpose, we have inserted unique cysteine residues at specific positions in the transmembrane and C-terminal region, using site-directed mutagenesis. Seven viable mutants with reasonable yield were harvested: A25C, V31C, T36C, G38C, T46C, A49C, and S50C. The mutant coat proteins were indistinguishable from wild type M13 coat protein with respect to their conformational and aggregational properties. The ESR data suggest that the amino acid positions 25 and 46 of the coat protein in DOPC vesicles are located close to the membrane-water interface. In this way the lysines at positions 40, 43, and 44 and the phenylalanines at positions 42 and 45 act as hydrophilic and hydrophobic anchors, respectively. The ESR spectra of site specific maleimido spin-labeled mutant coat proteins reconstituted into DOPC vesicles and solubilized in sodium cholate or SDS indicate that the local dynamics of the major coat protein is significantly affected by its structural environment...Continue Reading

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Citations

Aug 15, 2009·European Biophysics Journal : EBJ·Marcus A HemmingaDavid Stopar
Feb 1, 1990·Chemistry and Physics of Lipids·M Vauhkonen, P Somerharju
Nov 7, 1998·Biochimica Et Biophysica Acta·J A Killian
Sep 6, 2000·Trends in Biochemical Sciences·J A Killian, G von Heijne
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Apr 24, 2001·Protein Science : a Publication of the Protein Society·D BashtovyyT Páli
Jun 5, 2003·FEBS Letters·Erik Strandberg, J Antoinette Killian
Apr 20, 2006·Chemistry and Physics of Lipids·David StoparMarcus A Hemminga
Mar 28, 2003·Biochimica Et Biophysica Acta·David StoparMarcus A Hemminga
Aug 15, 2006·Biophysical Journal·David StoparMarcus A Hemminga
Jan 1, 2009·Journal of the History of Biology·Francisco Louçã
Jan 28, 1999·The Journal of Biological Chemistry·C G Bevans, A L Harris
Dec 22, 1999·The Journal of Biological Chemistry·A ArmulikS Johansson
Oct 28, 2003·Molecular Membrane Biology·Maurits R R de Planque, J Antoinette Killian
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Jul 4, 2007·Journal of Pharmaceutical and Biomedical Analysis·Salette ReisJosé L F C Lima
Nov 29, 2005·Journal of Chemical Information and Modeling·David StoparMarcus A Hemminga

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