The yeast Wsc1 cell surface sensor behaves like a nanospring in vivo.
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.
Dupres
Vincent
V
Unité de Chimie des Interfaces, Université Catholique de Louvain, Louvain-la-Neuve, Belgium.
Alsteens
David
D
Wilk
Sabrina
S
Hansen
Benjamin
B
Heinisch
Jürgen J
JJ
Dufrêne
Yves F
YF
D015374:Biosensing Techniques; D002462:Cell Membrane / Q000502:physiology* / Q000648:ultrastructure; D002473:Cell Wall / Q000502:physiology / Q000648:ultrastructure; D018914:Cells, Immobilized / Q000502:physiology / Q000648:ultrastructure; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000502:physiology; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology / Q000648:ultrastructure; D018625:Microscopy, Atomic Force / Q000379:methods*; D008958:Models, Molecular; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000502:physiology* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D012996:Solutions; D013314:Stress, Mechanical
D047908:Intracellular Signaling Peptides and Proteins; C091625:MID2 protein, S cerevisiae; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; C119383:SLG1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D012996:Solutions