A mass-spectrometric method for the estimation of the ratio of gamma-carboxyglutamic acid to glutamic acid at specific sites in proteins. Application to the N-terminal region of bovine prothrombin.

The Biochemical journal ·Vol. 187 ·No. 1 ·1980-04-01 ·Pages 239-43

Rose K, Priddle JD, Offord RE, Esnouf MP

Abstract

When a polypeptide containing gamma-carboxyglutamic acid is decarboxylated in 2H2O, residue of (gamma gamma-2H2)glutamic acid are formed. Subsequent proteolytic digestion produces peptides which contain at each site 2H2-substituted and unsubstituted glutamic acid in the same ratio as existed for gramma-carboxy-substitution. The peptides may be identified and this ratio determined by combined gas chromatography--mass spectrometry. We also discuss decarboxylation in 3H2O followed by amino-acid analysis and Edman degradation.

MeSH Terms

1-Carboxyglutamic Acid/analysis Amino Acid Sequence Animals Cattle Gas Chromatography-Mass Spectrometry Glutamates/analysis Methods Peptide Fragments/analysis Prothrombin

Chemicals

Glutamates Peptide Fragments 1-Carboxyglutamic Acid Prothrombin

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Rose K

Priddle J D

Offord R E

Esnouf M P

References (10)

10 references, click to expand

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[3H]diborane reduction of vitamin K-dependent calcium-binding proteins. Identification of a unique amino acid.
J Biol Chem. 1975 Apr 25;250(8):2968-72 PMID: 1123332
Isolation and characterization of vitamin K-dependent region of bovine blood clotting factor X.
Proc Natl Acad Sci U S A. 1975 Apr;72(4):1281-5 PMID: 1055402
A new carboxylation reaction. The vitamin K-dependent incorporation of H-14-CO3- into prothrombin.
J Biol Chem. 1975 Jun 25;250(12):4744-8 PMID: 1141226
A variant of prothrombin induced in cattle by prolonged administration of warfarin.
Biochim Biophys Acta. 1976 May 20;434(1):265-79 PMID: 938668
Mass-spectrometric identification and sequence location of the ten residues of the new amino acid (gamma-Carboxyglutamic acid) in the N-terminal region of prothrombin.
Biochem J. 1976 Mar 1;153(3):663-79 PMID: 942379
The separation and sequencing of permethylated peptides by mass spectrometry directly coupled to gas-liquid chromatography.
Biochem J. 1976 Sep 1;157(3):777-80 PMID: 985418
The gamma-carboxy glutamic acid content of human and bovine prothrombin following warfarin treatment.
Biochim Biophys Acta. 1977 Feb 22;490(2):471-6 PMID: 13857
The N-terminal sequences of blood coagulation factor X1 and X2 light chains. Mass-spectrometric identification of twelve residues of gamma-carboxyglutamic acid in their vitamin K-dependent domains.
Biochem J. 1978 Nov 1;175(2):613-27 PMID: 743214
Specific tritium labeling of gamma-carboxyglutamic acid in proteins.
Biochemistry. 1979 Oct 30;18(22):4992-9 PMID: 508728

Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj1870239

Published

1980-04-01

Pages

239-43

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1162513

Subset

External Links

PubMed source DOI: 10.1042/bj1870239 PMC Full Text View journal details

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A mass-spectrometric method for the estimation of the ratio of gamma-carboxyglutamic acid to glutamic acid at specific sites in proteins. Application to the N-terminal region of bovine prothrombin.

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