Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Biosensing Techniques*
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Cell Membrane / physiology*
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Cell Membrane / ultrastructure
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Cell Wall / physiology
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Cell Wall / ultrastructure
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Cells, Immobilized / physiology
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Cells, Immobilized / ultrastructure
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Intracellular Signaling Peptides and Proteins / chemistry
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Intracellular Signaling Peptides and Proteins / physiology
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Membrane Glycoproteins / chemistry
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Membrane Glycoproteins / physiology
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Membrane Proteins / physiology
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Membrane Proteins / ultrastructure
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Microscopy, Atomic Force / methods*
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Models, Molecular
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Protein Conformation
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Saccharomyces cerevisiae / physiology*
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Saccharomyces cerevisiae / ultrastructure
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / physiology
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Saccharomyces cerevisiae Proteins / ultrastructure
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Solutions
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Stress, Mechanical
Substances
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Intracellular Signaling Peptides and Proteins
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MID2 protein, S cerevisiae
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Membrane Glycoproteins
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Membrane Proteins
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SLG1 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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Solutions