Vitamin K-dependent carboxylase: increased efficiency of the carboxylation reaction.

Abstract:

:The in vitro vitamin K-dependent carboxylation of peptide- or protein-bound glutamate residues is generally studied in detergent-solubilized microsomes from rat or cow liver. Under the conditions usually employed, the efficiency of the carboxylation reaction is low (less than 1% of the carboxylatable residues is converted into gammacarboxyglutamate). Here we describe that this efficiency may be raised to 30% by carrying out the following adaptations: 1) carboxylase was purified about 100-fold from the solubilized microsomes, so that the enzyme was obtained in a highly concentrated form and could be added in excess: 2) the HCO-3 concentration in the reaction mixtures was raised to 50 mM and 3) a substrate was selected (decarboxylated osteocalcin from bovine bone) the Km of which had been shown to be low (10 microM) and it was added in rate-limiting amounts. Besides the fact that under these conditions the carboxylation reaction occurred with a higher efficiency than before, the adaptations also enabled us to express the carboxylation activity in terms of moles CO2 incorporated per mole of substrate.

journal_name

Thromb Haemost

authors

Soute BA,Ulrich MM,Vermeer C

subject

Has Abstract

pub_date

1987-02-03 00:00:00

pages

77-81

issue

1

eissn

0340-6245

issn

2567-689X

journal_volume

57

pub_type

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