Document Detail


Quantitative identification of N-terminal amino acids in proteins by radiolabeled reductive methylation and amino acid analysis: application to human erythrocyte acetylcholinesterase.
MedLine Citation:
PMID:  4037298     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A novel method of determining N-terminal amino acids in proteins is introduced. Reductive methylation of a protein with radiolabeled formaldehyde methylates both the alpha-amino group of the N-terminal amino acid and the epsilon-amino groups of Lys residues. The radiomethylated amino acids are stable to acid hydrolysis, and each of 16 possible hydrolysis-stable N-terminal amino acids can be identified by the unique elution positions of its N alpha-methyl and N alpha,N alpha-dimethyl derivatives with an appropriate amino acid analyzer elution schedule. The technique is at least as sensitive as other N-terminal amino acid determinations and, in addition, permits a quantitative evaluation of the number of N-terminal groups in a sample. Reductive methylation of bovine serum albumin revealed N-terminal Asp at a stoichiometry of 0.97 amino acid residue per polypeptide, while methylation of prolactin resulted in 0.86 residue of N-terminal Thr per polypeptide. Human erythrocyte acetylcholinesterase contained two N-terminal amino acids with stoichiometries of 0.66 Glu and 0.34 Arg per 70-kDa subunit. Identification of Glu as the principal N-terminus of acetylcholinesterase was confirmed by Edman sequencing.
Authors:
R Haas; T L Rosenberry
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  148     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1985 Jul 
Date Detail:
Created Date:  1985-10-10     Completed Date:  1985-10-10     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  154-62     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Acetylcholinesterase / blood*
Amino Acid Sequence
Amino Acids / analysis*
Borohydrides
Formaldehyde
Humans
Methylation*
Oxidation-Reduction
Grant Support
ID/Acronym/Agency:
NS-07118/NS/NINDS NIH HHS; NS-16577/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Borohydrides; 25895-60-7/sodium cyanoborohydride; 50-00-0/Formaldehyde; EC 3.1.1.7/Acetylcholinesterase

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


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