gamma-Glutamate and beta-hydroxyaspartate in proteins.
Vitamin K-dependent coagulation plasma proteins possess from 9-12 residues of gamma-carboxyglutamic acid (Gla) distributed over a ca. 45 amino acid peptide sequence, i.e., the Gla domain, which encompasses the NH2-terminal region. In addition, epidermal growth factor (EGF) homology units present in many of these same proteins contain beta-hydroxyaspartate (Hya) residues, which is a modification decoupled from gamma-carboxylation. The function of Gla residues in these proteins, viz., prothrombin, coagulation factors VII, IX, and X, along with anticoagulant protein C and protein S, is to coordinate Ca2+. This results in a large conformational alteration in the proteins or peptides, which allows adsorption to membrane phospholipids (PL), an event that is critical is to their proper functions in the blood coagulation system. Less certain is the role of Hya in EGF domains, but it has been proposed that modification at this residue may negatively regulate fucosylation of these regions. In several proteins, these modules also interact with Ca2+, but it has been shown that although the particular aspartate containing the beta-OH group is critical to that interaction, beta-hydroxylation of that Asp residue is not. Because of their widespread distribution, quantitative detection protocols for both Gla and Hya are of importance. It is the purpose of this communication to detail a reliable method for these analyses that is employed in our laboratories.
Castellino
Francis J
FJ
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA.
Ploplis
Victoria A
VA
Zhang
Li
L
D000327:Adsorption; D000818:Animals; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D018698:Glutamic Acid / Q000032:analysis*; D006801:Humans; D008563:Membrane Lipids / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*
D008563:Membrane Lipids; D010743:Phospholipids; D011506:Proteins; C011274:3-hydroxyaspartic acid; D001224:Aspartic Acid; D018698:Glutamic Acid