Document Detail

A pulse-chase-competition experiment to determine if a folding intermediate is on or off-pathway: application to ribonuclease A.
MedLine Citation:
PMID:  9784375     Owner:  NLM     Status:  MEDLINE    
A modified pulse-chase experiment is applied to determine if the native-like intermediate IN of ribonuclease A is on or off-pathway. The 1H label retained in the native protein is compared when separate samples of 1H-labeled IN and unfolded protein are allowed to fold to native in identical conditions. The solvent is 2H2O and the pH* is such that the unfolded protein rapidly exchanges its peptide NH protons with solvent, and IN does not. If IN is on-pathway, more 1H-label will be retained in the test sample starting with IN than in the control sample starting with unfolded protein. The results show that IN is a productive (on-pathway) intermediate. Application of the modified pulse-chase experiment to the study of rapidly formed folding intermediates may be possible when a rapid mixing device is used.
D V Laurents; M Bruix; M Jamin; R L Baldwin
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of molecular biology     Volume:  283     ISSN:  0022-2836     ISO Abbreviation:  J. Mol. Biol.     Publication Date:  1998 Oct 
Date Detail:
Created Date:  1998-11-20     Completed Date:  1998-11-20     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985088R     Medline TA:  J Mol Biol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  669-78     Citation Subset:  IM    
Copyright Information:
Copyright 1998 Academic Press.
Department of Biochemistry, Beckman Center, Stanford, CA, 94305-5307, USA.
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MeSH Terms
Hydrogen-Ion Concentration
Protein Denaturation
Protein Folding*
Ribonuclease, Pancreatic / chemistry*
Grant Support
Reg. No./Substance:
0/Protons; EC, Pancreatic

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