Document Detail


Complete amino acid analysis of proteins from a single hydrolysate.
MedLine Citation:
PMID:  178649     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
An analytical procedure which affords the precise amino acid composition of a protein or a peptide from a single hydrolysate is described. This method utilizes 4 N methanesulfonic acid containing 0.2% 3-(2-aminoethyl)indole, rather then 6N HCl as a catalyst for hydrolysis. The hydrolysis is carried out in vacuo (20 mu) at 115 degrees for 22 to 72 hours. Half-cystine is determined as S-sulfocysteine by treating the hydrolysate with dithiothreitol followed by an excess of tetrathionate. The values of all amino acids, including tryptophan and half-cystine, were close to the expected theoretical values for the proteins examined. The method has the advantage that the neutralized hydrolysate can be applied directly to an ion exchange column. Further, the method is capable of distinguishing between free sulfhydryl groups as S-carbosymethylcysteine and disulfides as S-sulfocysteine. A limitation of the procedure is that tryptophan remains sensitive to the presence of carbohydrate in the sample.
Authors:
R J Simpson; M R Neuberger; T Y Liu
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  251     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1976 Apr 
Date Detail:
Created Date:  1976-08-02     Completed Date:  1976-08-02     Revised Date:  2009-10-27    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1936-40     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / analysis*
Binding Sites
Chemical Phenomena
Chemistry
Chromatography, Ion Exchange / methods
Endopeptidases*
Ficain*
Glyceraldehyde-3-Phosphate Dehydrogenases*
Kinetics
Mesylates
Papain*
Protein Binding
Protein Hydrolysates*
Temperature
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Mesylates; 0/Protein Hydrolysates; EC 1.2.1.-/Glyceraldehyde-3-Phosphate Dehydrogenases; EC 3.4.-/Endopeptidases; EC 3.4.22.2/Papain; EC 3.4.22.3/Ficain

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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