Lipid and signal peptide-induced conformational changes within the C-domain of Escherichia coli SecA protein

Biochemistry
H DingD B Oliver

Abstract

SecA ATPase is an essential component of the Sec-dependent protein translocation machinery. Upon interaction with the plasma membrane containing SecYE, preprotein, and ATP, SecA undergoes cycles of membrane insertion and retraction resulting in the translocation of segments of the preprotein to the trans side of the membrane. To study the structural basis of SecA function, we employed fluorescence spectroscopy along with collisional quenchers with a set of SecA proteins containing single tryptophan substitutions. Our data show that among the seven naturally occurring tryptophan residues of Escherichia coli SecA, only the three tryptophan residues contained within the C-domain contributed significantly to the fluorescence signal, and they occupied distinct local environments in solution: Trp723 and Trp775 were found to be relatively solvent accessible and inaccessible, respectively, while Trp701 displayed an intermediate level of solvent exposure. Exposure to increased temperature or interaction with model membranes or signal peptide elicited a similar conformational response from SecA based upon the fluorescence signals of the SecA-W775F and SecA-W723F mutant proteins. Specifically, Trp775 became more solvent exposed, while Trp...Continue Reading

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Citations

Apr 7, 2005·Journal of Molecular Biology·Linda L RandallSimon J S Hardy
Feb 25, 2003·Chemistry and Physics of Lipids·Luís M S LouraManuel Prieto
Oct 19, 2005·Biochemistry·Monika Musial-SiwekDebra A Kendall
Nov 22, 2012·Journal of the American Chemical Society·Dorothy M KimJohn F Hunt
Jan 13, 2006·Nucleic Acids Research·Mary K Smalley, Scott K Silverman
Jul 17, 2004·Proceedings of the National Academy of Sciences of the United States of America·Andrew R OsborneTom A Rapoport
Oct 6, 2007·Biopolymers·Eugenia M CléricoLila M Gierasch
Oct 3, 2006·Biochimica Et Biophysica Acta·Sharyn L Rusch, Debra A Kendall
Jun 8, 2006·Journal of Biochemistry and Molecular Biology·Ji Yeun ShinTaeho Ahn
Nov 18, 2004·Biochimica Et Biophysica Acta·Eleftheria Vrontou, Anastassios Economou
Oct 31, 2002·The Journal of Biological Chemistry·Jordi BenachJohn F Hunt
Oct 28, 2017·International Journal of Molecular Sciences·Joy M Cote, Erika A Taylor
Oct 26, 2005·The Journal of Biological Chemistry·Efrosyni PapanikouAnastassios Economou
Apr 11, 2019·Protein Science : a Publication of the Protein Society·Guillaume RousselStephen H White
Feb 5, 2002·The Journal of Biological Chemistry·Catherine BaudAnastassios Economou
Mar 10, 2004·The Journal of Biological Chemistry·Eleftheria VrontouAnastassios Economou

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