Nature of the vitamin K-dependent CO2 fixation in microsomal membranes.
Vitamin K is a component of a membrane-bound enzyme complex which catalyzes the posttranslational carboxylation of peptide-bound glutamate to form the gamma-carboxyglutamate (Gla) residues of prothrombin. The reaction requires reduced vitamin K, bicarbonate, oxygen, and a carboxylase, and does not require ATP. In a Triton X-100 solubilized carboxylase system, it was found that the naphthoquinone ring structure is essential for activity, as is the 2-methyl group. Menaquinone homologs from MK-1 to MK-4 all had carboxylase activity, whereas menadione was inactive. However, dithiothreitol and other thiols form thioethers with menadione, which restores considerable carboxylation activity to the provitamin. Hydrogenation of the beta-gamma double bond in phylloquinone reduced its activity only slightly. The active species of "CO2" utilized in this carboxylation is CO2 and not bicarbonate. Ribosomes contain Gla residues and are labeled with CO2 when whole microsomes are incubated with CO2 in the presence of NADH and vitamin K. About 25% of the activity is releasable with puromycin, suggesting that Gla residues are formed on both the nascent chains and the structural proteins of ribosomes. The deoxycholate-solubilized carboxylase system can be dialyzed to yield membranous vesicles with enhanced carboxylase activity. The warfarin-binding protein from normal rats, but not that from warfarin-resistant rats, further enhances the carboxylase activity of these reformed vesicles.
D015055:1-Carboxyglutamic Acid / Q000378:metabolism*; D000818:Animals; D002245:Carbon Dioxide / Q000378:metabolism*; D002262:Carboxy-Lyases / Q000378:metabolism; D002352:Carrier Proteins / Q000494:pharmacology; D005971:Glutamates / Q000378:metabolism*; D007425:Intracellular Membranes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D051381:Rats; D012269:Ribosomal Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D014812:Vitamin K / Q000378:metabolism*; D014813:Vitamin K Deficiency / Q000378:metabolism; D014859:Warfarin / Q000494:pharmacology
D002352:Carrier Proteins; D005971:Glutamates; D010455:Peptides; D012269:Ribosomal Proteins; D014812:Vitamin K; D002245:Carbon Dioxide; D015055:1-Carboxyglutamic Acid; D014859:Warfarin; D002262:Carboxy-Lyases