Document Detail


Dynamic properties of membrane proteins: reversible insertion into membrane vesicles of a colicin E1 channel-forming peptide.
MedLine Citation:
PMID:  2459708     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The binding of colicin E1 and its COOH-terminal channel-forming peptides to artificial membrane vesicles has an optimum at acidic pH values. The Mr 18,000 thermolytic peptide inserted into membrane vesicles at pH 4.0 has a limited accessibility to exogenous protease. It is converted by trypsin cleavage after Lys-381 and Lys-382 to a lower Mr 14,000 peptide. However, when the pH of a vesicle suspension to which peptide has been bound at pH 4.0 is shifted to 6.0, the accessibility to protease increased greatly. This was shown (i) by the large decrease in the amount of Mr 14,000 or Mr 18,000 peptide after the pH 4----6 shift and treatment with trypsin or Pronase, consistent with (ii) a previously observed decrease in membrane-bound radiolabeled peptide after protease treatment. (iii) When a photoactivable nitrene-generating phospholipid probe was used to label the colicin peptide inserted into the bilayer, the extent of labeling decreased by a factor of 3 when the pH was shifted from 4.0 to 6.5. (iv) Colicin peptide added to vesicles at pH 4.0 can "hop" to other vesicles if the pH and ionic strength of donor vesicles are successively increased. It is proposed that deprotonation of acidic residues in contact with the hydrophobic bilayer or the membrane surface destabilizes the inserted channel and causes it to be extruded from the membrane. The pH-dependent extrusion of the inserted colicin channel provides an example of dynamic properties of an intrinsic membrane protein.
Authors:
S Xu; W A Cramer; A A Peterson; M Hermodson; C Montecucco
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  85     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1988 Oct 
Date Detail:
Created Date:  1988-11-21     Completed Date:  1988-11-21     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  7531-5     Citation Subset:  IM    
Affiliation:
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Chromatography, High Pressure Liquid
Colicins / metabolism*
Hydrogen-Ion Concentration
Ion Channels / metabolism*
Membrane Proteins / metabolism*
Membranes, Artificial*
Molecular Sequence Data
Peptide Fragments / metabolism*
Grant Support
ID/Acronym/Agency:
GM-18457/GM/NIGMS NIH HHS; GM-24602/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Colicins; 0/Ion Channels; 0/Membrane Proteins; 0/Membranes, Artificial; 0/Peptide Fragments
Comments/Corrections

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