The yeast Wsc1 cell surface sensor behaves like a nanospring in vivo

被引:113
作者
Dupres, Vincent [1 ]
Alsteens, David [1 ]
Wilk, Sabrina [2 ]
Hansen, Benjamin [2 ]
Heinisch, Juergen J. [2 ]
Dufrene, Yves F. [1 ]
机构
[1] Catholic Univ Louvain, Unite Chim Interfaces, B-3000 Louvain, Belgium
[2] Univ Osnabruck, Fachbereich Biol Chem, AG Genet, D-4500 Osnabruck, Germany
关键词
ATOMIC-FORCE MICROSCOPY; SINGLE-MOLECULAR RECOGNITION; SACCHAROMYCES-CEREVISIAE; INTEGRITY; PROTEIN; RECEPTOR; DOMAINS; MID2; SPECTROSCOPY; PATHWAYS;
D O I
10.1038/nchembio.220
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Here we report on in vivo measurement of the mechanical behavior of a cell surface sensor using single-molecule atomic force microscopy. We focus on the yeast wall stress component sensor Wsc1, a plasma membrane protein that is thought to function as a rigid probe of the cell wall status. We first map the distribution of individual histidine-tagged sensors on living yeast cells by scanning the cell surface with atomic force microscopy tips carrying nitrilotriacetate groups. We then show that Wsc1 behaves like a linear nanospring that is capable of resisting high mechanical force and of responding to cell surface stress. Both a genomic pmt4 deletion and the insertion of a stretch of glycines in Wsc1 result in substantial alterations in protein spring properties, supporting the important role of glycosylation at the extracellular serine/threonine-rich region.
引用
收藏
页码:857 / 862
页数:6
相关论文
共 36 条
[1]   Real-time imaging of the surface topography of living yeast cells by atomic force microscopy [J].
Ahimou, FO ;
Touhami, A ;
Dufrêne, YF .
YEAST, 2003, 20 (01) :25-30
[2]  
ARVANITIDIS A, 1994, J BIOL CHEM, V269, P8911
[3]   Molecular nanosprings in spider capture-silk threads [J].
Becker, N ;
Oroudjev, E ;
Mutz, S ;
Cleveland, JP ;
Hansma, PK ;
Hayashi, CY ;
Makarov, DE ;
Hansma, HG .
NATURE MATERIALS, 2003, 2 (04) :278-283
[4]   Functional links between membrane transport and the spectrin cytoskeleton [J].
Dubreuil, Ronald R. .
JOURNAL OF MEMBRANE BIOLOGY, 2006, 211 (03) :151-161
[5]   Force-clamp spectroscopy monitors the folding trajectory of a single protein [J].
Fernandez, JM ;
Li, HB .
SCIENCE, 2004, 303 (5664) :1674-1678
[6]   NEW YEAST-ESCHERICHIA-COLI SHUTTLE VECTORS CONSTRUCTED WITH INVITRO MUTAGENIZED YEAST GENES LACKING 6-BASE PAIR RESTRICTION SITES [J].
GIETZ, RD ;
SUGINO, A .
GENE, 1988, 74 (02) :527-534
[7]   Baker's yeast as a tool for the development of antifungal drugs which target cell integrity - an update [J].
Heinisch, Juergen J. .
EXPERT OPINION ON DRUG DISCOVERY, 2008, 3 (08) :931-943
[8]   Detection and localization of single molecular recognition events using atomic force microscopy [J].
Hinterdorfer, P ;
Dufrêne, YF .
NATURE METHODS, 2006, 3 (05) :347-355
[9]   WHY ARE PROTEINS O-GLYCOSYLATED [J].
JENTOFT, N .
TRENDS IN BIOCHEMICAL SCIENCES, 1990, 15 (08) :291-294
[10]   Mechanical strain regulates syndecan-4 expression and shedding in smooth muscle cells through differential activation of MAP kinase signaling pathways [J].
Julien, Matheau A. ;
Wang, Peiyi ;
Haller, Carolyn A. ;
Wen, Jing ;
Chaikof, Elliot L. .
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2007, 292 (01) :C517-C525