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Processive post-translational modification. Vitamin K-dependent carboxylation of a peptide substrate.

Publication ,  Journal Article
Morris, DP; Stevens, RD; Wright, DJ; Stafford, DW
Published in: J Biol Chem
December 22, 1995

Mass spectrometry has been used to demonstrate that vitamin K-dependent carboxylation is a processive post-translational modification (i.e. multiple carboxylations occur during a single association between enzyme and substrate). Purified vitamin K-dependent carboxylase can carboxylate as many as 12 glutamate residues in FIXQ/S, a peptide substrate based on amino acids -18 to 41 of the human blood clotting enzyme factor IX. Mass spectrometry was used to determine the number of gamma-carboxyl groups added to FIXQ/S by the carboxylase during an in vitro reaction. Despite the fact that most substrate molecules in a reaction were uncarboxylated, almost all carboxylated FIXQ/S molecules were carboxylated many times. This observation can only be explained by two types of mechanisms. In a processive mechanism, multiple carboxylations could occur during a single substrate binding event. Alternatively, a distributive mechanism could result in the observed behavior if the initial carboxylation event results in a substrate that is additionally carboxylated far more efficiently than the uncarboxylated FIXQ/S. Kinetic experiments and arguments were used to show that the vitamin K-dependent carboxylase is not distributive but rather is one of the first well documented examples of an enzyme that catalyzes a processive post-translation modification.

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Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

December 22, 1995

Volume

270

Issue

51

Start / End Page

30491 / 30498

Location

United States

Related Subject Headings

  • Substrate Specificity
  • Recombinant Proteins
  • Protein Processing, Post-Translational
  • Peptides
  • Peptide Fragments
  • Peptide Biosynthesis
  • Molecular Sequence Data
  • Mass Spectrometry
  • Ligases
  • Kinetics
 

Citation

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Morris, D. P., Stevens, R. D., Wright, D. J., & Stafford, D. W. (1995). Processive post-translational modification. Vitamin K-dependent carboxylation of a peptide substrate. J Biol Chem, 270(51), 30491–30498. https://doi.org/10.1074/jbc.270.51.30491
Morris, D. P., R. D. Stevens, D. J. Wright, and D. W. Stafford. “Processive post-translational modification. Vitamin K-dependent carboxylation of a peptide substrate.J Biol Chem 270, no. 51 (December 22, 1995): 30491–98. https://doi.org/10.1074/jbc.270.51.30491.
Morris DP, Stevens RD, Wright DJ, Stafford DW. Processive post-translational modification. Vitamin K-dependent carboxylation of a peptide substrate. J Biol Chem. 1995 Dec 22;270(51):30491–8.
Morris, D. P., et al. “Processive post-translational modification. Vitamin K-dependent carboxylation of a peptide substrate.J Biol Chem, vol. 270, no. 51, Dec. 1995, pp. 30491–98. Pubmed, doi:10.1074/jbc.270.51.30491.
Morris DP, Stevens RD, Wright DJ, Stafford DW. Processive post-translational modification. Vitamin K-dependent carboxylation of a peptide substrate. J Biol Chem. 1995 Dec 22;270(51):30491–30498.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

December 22, 1995

Volume

270

Issue

51

Start / End Page

30491 / 30498

Location

United States

Related Subject Headings

  • Substrate Specificity
  • Recombinant Proteins
  • Protein Processing, Post-Translational
  • Peptides
  • Peptide Fragments
  • Peptide Biosynthesis
  • Molecular Sequence Data
  • Mass Spectrometry
  • Ligases
  • Kinetics