Document Detail


Unfolding dynamics of the mucin SEA domain probed by force spectroscopy suggest that it acts as a cell-protective device.
MedLine Citation:
PMID:  23331320     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
MUC1 and other membrane-associated mucins harbor long, up to 1 μm, extended highly glycosylated mucin domains and sea urchin sperm protein, enterokinase and agrin (SEA) domains situated on their extracellular parts. These mucins line luminal tracts and organs, and are anchored to the apical cell membrane by a transmembrane domain. The SEA domain is highly conserved and undergoes a molecular strain-dependent autocatalytic cleavage during folding in the endoplasmic reticulum, a process required for apical plasma membrane expression. To date, no specific function has been designated for the SEA domain. Here, we constructed a recombinant protein consisting of three SEA domains in tandem and used force spectroscopy to assess the dissociation force required to unfold individual, folded SEA domains. Force-distance curves revealed three peaks, each representing unfolding of a single SEA domain. Fitting the observed unfolding events to a worm-like chain model yielded an average contour length of 32 nm per SEA domain. Analysis of forces applied on the recombinant protein revealed an average unfolding force of 168 pN for each SEA domain at a loading rate of 25 nN·s(-1). Thus, the SEA domain may act as a breaking point that can dissociate before the plasma membrane is breached when mechanical forces are applied to cell surfaces.
Authors:
Thaher Pelaseyed; Michael Zäch; Asa C Petersson; Frida Svensson; Denny G A Johansson; Gunnar C Hansson
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2013-02-21
Journal Detail:
Title:  The FEBS journal     Volume:  280     ISSN:  1742-4658     ISO Abbreviation:  FEBS J.     Publication Date:  2013 Mar 
Date Detail:
Created Date:  2013-03-15     Completed Date:  2013-05-02     Revised Date:  2014-03-06    
Medline Journal Info:
Nlm Unique ID:  101229646     Medline TA:  FEBS J     Country:  England    
Other Details:
Languages:  eng     Pagination:  1491-501     Citation Subset:  IM    
Copyright Information:
© 2013 The Authors Journal compilation © 2013 FEBS.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biomechanical Phenomena
CHO Cells
Cell Membrane / chemistry
Cricetinae
Enzyme-Linked Immunosorbent Assay
Microscopy, Atomic Force / methods*
Models, Molecular
Mucin-1 / chemistry*,  genetics
Mutagenesis, Site-Directed
Protein Conformation
Protein Stability
Protein Structure, Tertiary
Protein Unfolding*
Proteolysis
Recombinant Proteins / chemistry,  genetics
Stress, Mechanical
Temperature
Transfection
Grant Support
ID/Acronym/Agency:
U01 AI095473/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Mucin-1; 0/Recombinant Proteins
Comments/Corrections

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