Document Detail


Isolation and characterization of the reaction product of 4-diazobenzenesulfonic acid and gamma-carboxyglutamic acid: modification of the assay for measurement of beta-carboxyaspartic acid.
MedLine Citation:
PMID:  8135347     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
gamma-Carboxyglutamic acid (Gla)-containing proteins are extracellular proteins with enhanced cation or mineral-binding properties. Discovery of new Gla-containing proteins is facilitated by methods that decrease the number of steps and time involved in assaying for Gla. Reaction of 4-diazobenzenesulfonic acid (DBS) and Gla or Gla-containing proteins produces an intensely red-colored product. This method has been used to identify Gla-containing proteins in crude extracts of proteins. The reaction product of Gla and DBS has been purified by reversed-phase HPLC and characterized by uv-visible spectroscopy and electrospray ionization (ESI) mass spectrometry (MS). The red-colored product exhibits an absorption maximum at 530 nm. The ESI-MS data of the colored derivative of Gla are consistent with replacement of the two gamma-carboxyl groups in Gla with two DBS groups. DBS also reacts with another malonic acid derivative, beta-carboxyaspartic acid (Asa). The optimum conditions for colorimetric assay of Asa were established, and the potential of this reaction as an assay for Asa and Asa-containing proteins was studied.
Authors:
S K Nishimoto; J Zhao; C Dass
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  216     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1994 Jan 
Date Detail:
Created Date:  1994-04-18     Completed Date:  1994-04-18     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  159-64     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, University of Tennessee, Memphis 38163.
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MeSH Terms
Descriptor/Qualifier:
1-Carboxyglutamic Acid*
Aspartic Acid / analogs & derivatives*,  analysis
Chromatography, High Pressure Liquid
Colorimetry
Diazonium Compounds*
Mass Spectrometry
Sulfanilic Acids*
Grant Support
ID/Acronym/Agency:
AR35056/AR/NIAMS NIH HHS; NS28025/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Diazonium Compounds; 0/Sulfanilic Acids; 2154-66-7/diazobenzenesulfonic acid; 53445-96-8/1-Carboxyglutamic Acid; 56-84-8/Aspartic Acid; 75898-26-9/beta-carboxyaspartic acid

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


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