PMID: 8962080Dec 10, 1996Paper

A general method for determining helix packing in membrane proteins in situ: helices I and II are close to helix VII in the lactose permease of Escherichia coli

Proceedings of the National Academy of Sciences of the United States of America
J Wu, H R Kaback

Abstract

It was previously shown that coexpression of the lactose permease of Escherichia coli in two contiguous fragments leads to functional complementation. We demonstrate here that site-directed thiol crosslinking of coexpressed permease fragments can be used to determine helix proximity in situ without the necessity of purifying the permease. After coexpression of the six N-terminal (N6) and six C-terminal (C6) transmembrane helices, each with a single Cys residue, crosslinking was carried out in native membranes and assessed by the mobility of anti-C-terminal-reactive polypeptides on immunoblots. A Cys residue at position 242 or 245 (helix VII) forms a disulfide with a Cys residue at either position 28 or 29 (helix I), but not with a Cys residue at position 27, which is on the opposite face of helix I, thereby indicating that the face of helix I containing Pro-28 and Phe-29 is close to helix VII. Similarly, a Cys residue at position 242 or 245 (helix VII) forms a disulfide with a Cys residue at either position 52 or 53 (helix II), but not with a Cys residue at position 54. Furthermore, low-efficiency crosslinking is observed between a Cys residue at position 52 or 53 and a Cys residue at position 361 (helix XI). The results indica...Continue Reading

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Citations

Jul 1, 1997·Protein Science : a Publication of the Protein Society·J SunH R Kaback
Aug 1, 1997·Current Opinion in Structural Biology·H R KabackJ Wu
Aug 3, 2001·Nature Reviews. Molecular Cell Biology·H R KabackA B Weinglass
Mar 6, 2008·Proceedings of the National Academy of Sciences of the United States of America·Yonggang ZhouH Ronald Kaback
Feb 5, 2009·Proceedings of the National Academy of Sciences of the United States of America·P BarthD Baker
Mar 20, 2009·Proceedings of the National Academy of Sciences of the United States of America·Giani OanceaRobert Tampé
Jun 27, 2013·Proceedings of the National Academy of Sciences of the United States of America·Mousheng WuLei Zheng
Oct 20, 2009·The Journal of Biological Chemistry·Zhen TaoGary Rudnick
Jul 7, 2011·The Journal of Biological Chemistry·Yonggang ZhouH Ronald Kaback
Feb 20, 2003·Journal of Bacteriology·Teruhisa HiraiSriram Subramaniam
Oct 14, 2000·Annual Review of Cell and Developmental Biology·R E Dalbey, A Kuhn
Oct 23, 2002·Proceedings of the National Academy of Sciences of the United States of America·Paul L SorgenMark E Girvin
May 27, 1997·Proceedings of the National Academy of Sciences of the United States of America·H R Kaback
Aug 30, 2001·Proceedings of the National Academy of Sciences of the United States of America·H HastrupJ A Javitch
Feb 17, 1999·Proceedings of the National Academy of Sciences of the United States of America·Y Zhang, B I Kanner
Jan 25, 2006·British Journal of Pharmacology·Mariana Oana Popa, Holger Lerche
May 30, 2006·Protein Science : a Publication of the Protein Society·Richard B JacobsenJoseph S Schoeniger
Jan 2, 2010·Journal of Molecular Biology·Zhenyu LiuH Ronald Kaback
Jul 30, 2004·Biochimica Et Biophysica Acta·E PadanL Galili
Nov 30, 2018·The Journal of Biological Chemistry·Sujeet KumarNatividad Ruiz
Aug 11, 2005·The Journal of Biological Chemistry·Amal K Bera, Myles H Akabas
Apr 22, 2015·Nature Communications·Kazuo TatebayashiHaruo Saito
Aug 14, 1999·The Journal of Biological Chemistry·E H MantingA J Driessen
Dec 24, 2002·The Journal of Biological Chemistry·Wen GuoJonathan A Javitch
Feb 19, 2003·The Journal of Biological Chemistry·Shushi NagamoriH Ronald Kaback
Jan 11, 2007·The Biochemical Journal·Emily R SlepkovLarry Fliegel
Oct 8, 1998·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·S FrillingosH R Kaback
Nov 12, 2003·The Journal of Biological Chemistry·Thomas L KashNeil L Harrison
Dec 14, 2006·The Journal of Biological Chemistry·Meike HergetRupert Abele
Nov 26, 1997·Biochimica Et Biophysica Acta·A C WeissbornT H Wilson

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