Document Detail

Analysis of 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin amino acids at sub-picomole levels by high-performance liquid chromatography: simultaneous manual sequencing of picomole quantities of several polypeptides.
MedLine Citation:
PMID:  2500874     Owner:  NLM     Status:  MEDLINE    
A single-column high-performance liquid chromatographic separation of 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin amino acid derivatives, generated during polypeptide sequence analysis by the 4-N,N-dimethylaminoazobenzene 4'-isothiocyanate/phenylisothiocyanate double coupling technique, is described. Recovery of the serine and threonine derivatives was improved by substituting boron trifluoride-diethyl etherate for trifluoroacetic acid in the thiazolinone cleavage reactions. Residues, including the S-carboxymethyl derivative of cysteine, were assigned after a single injection and a cycle time of 30 min. Quantities of 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin amino acid derivatives as low as 100 fmol were detected. Interference of sequencing artefacts with residue assignment was avoided. This technique allows simultaneous manual sequencing of several proteins or peptides at the level of a few picomoles.
P M Fischer; M E Howden
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical biochemistry     Volume:  177     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1989 Feb 
Date Detail:
Created Date:  1989-08-10     Completed Date:  1989-08-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  46-9     Citation Subset:  IM    
Division of Biological and Health Sciences, Deakin University, Geelong, Victoria, Australia.
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MeSH Terms
Amino Acid Sequence*
Amino Acids / analysis*
Chromatography, High Pressure Liquid
Hydantoins / analysis*
Thiohydantoins / analysis*
p-Dimethylaminoazobenzene / analogs & derivatives*,  analysis
Reg. No./Substance:
0/Amino Acids; 0/Hydantoins; 0/Isothiocyanates; 0/Thiocyanates; 0/Thiohydantoins; 103-72-0/phenylisothiocyanate; 60-11-7/p-Dimethylaminoazobenzene; 72683-57-9/DABTH; 7612-98-8/4-(N,N-dimethylaminoazobenzene)-4'-isothiocyanate

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