Intracellular maturation of the gamma-carboxyglutamic acid (Gla) region in prothrombin coincides with release of the propeptide.
Vitamin K-dependent coagulation factors undergo several post-translational modifications before the proteins are secreted into the blood as functional zymogens of the coagulation system. The modifications include Asn-linked glycosylation, Asn/Asp hydroxylation, removal of a signal peptide for translocation of the polypeptide into the endoplasmic reticulum and removal of a propeptide which, when attached to the intracellular coagulation factor precursor, directs the protein for vitamin K-dependent gamma-carboxylation. gamma-Carboxylation of targeted Glu residues results in formation of Ca(2+)-binding gamma-carboxyglutamic acid (Gla) residues. Ca2+ binding by these residues induces a conformational change in the protein which is a necessary event for optimal activation or activity of the clotting factor in blood. In the present study we have monitored the intracellular prothrombin precursor in the secretory pathway of liver cells to determine the effect that the propeptide has on Ca(2+)-dependent folding of the protein. The data provide evidence that the Ca(2+)-induced conformational change required for activation of prothrombin coincides with release of the propeptide in the trans-Golgi apparatus of the liver cell and elucidates an important function for the endoproteinase furin in biosynthesis of vitamin K-dependent clotting factors.
Wallin
R
R
Department of Medicine, Bowman Gray School of Medicine, Winston-Salem, NC 27517-1058.
Stanton
C
C
Hutson
S M
SM
D015055:1-Carboxyglutamic Acid / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D006056:Golgi Apparatus / Q000378:metabolism; D015151:Immunoblotting; D008099:Liver / Q000648:ultrastructure; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D017510:Protein Folding; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011516:Prothrombin / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D011498:Protein Precursors; D015055:1-Carboxyglutamic Acid; D011516:Prothrombin; D002118:Calcium