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Related Experiment Videos

Processing of prothrombin in the secretory pathway.

C Stanton1, R Taylor, R Wallin

  • 1Department of Medicine, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC 27103.

The Biochemical Journal
|July 1, 1991
PubMed
Summary

Prothrombin precursors undergo modifications in the Golgi apparatus, increasing in mass. The propeptide is removed by a specific enzyme, and vitamin K-dependent carboxylation occurs, suggesting gamma-carboxyglutamic acid synthesis late in processing.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Prothrombin is a key protein in blood coagulation.
  • Its synthesis involves post-translational modifications.
  • Understanding these modifications is crucial for hemostasis research.

Purpose of the Study:

  • To identify and characterize prothrombin precursors during synthesis.
  • To investigate the role of the propeptide in prothrombin processing.
  • To elucidate the location and substrate specificity of vitamin K-dependent carboxylase.

Main Methods:

  • Antibody-based identification of prothrombin precursors in cellular compartments.
  • Analysis of molecular mass changes during processing.
  • Assay of vitamin K-dependent carboxylase activity in Golgi fractions.

Related Experiment Videos

  • Use of synthetic peptides as carboxylase substrates.

    • Main Results:

    • Prothrombin precursors are modified in the Golgi apparatus, increasing from 78 kDa to 83 kDa.
    • The propeptide is part of the precursor during Golgi modifications and is cleaved by a Ca(2+)-dependent serine proteinase.
    • Vitamin K-dependent carboxylase activity is highest in cis-Golgi cisternae.
    • Different peptide regions of prothrombin are preferred substrates for the carboxylase in microsomes versus Golgi.
    • Prothrombin precursor gains negative charges in the Golgi, indicative of gamma-carboxyglutamic acid (Gla) residue synthesis.

    Conclusions:

    • The propeptide is integral to prothrombin processing within the Golgi apparatus.
    • Prothrombin maturation involves proteolytic cleavage and vitamin K-dependent carboxylation in the Golgi.
    • Gamma-carboxyglutamic acid residues are likely synthesized late in the secretory pathway.