title: gamma-Carboxyglutamic acid content of hepatocellular carcinoma-associated des-gamma-carboxy prothrombin.
abstract: Serum des-gamma-carboxy prothrombin (DCP) is a useful marker for the diagnosis of hepatocellular carcinoma (HCC), but the exact mechanism of its synthesis and its structural properties in liver diseases are unknown. DCP is measured by the monoclonal antibody MU-3. The purpose of this study was to examine the epitope of MU-3 and to characterize the differences in DCP between HCC and benign liver diseases. The epitope of MU-3 was examined by ELISA using prothrombin Gla domain polypeptides and was determined to be amino acid residues 17-27 of the prothrombin Gla domain, which has four gamma-carboxyglutamic acid residues (Gla) at positions 19, 20, 25 and 26. Peptides having a glutamic acid residue (Glu) at these positions reacted strongly to MU-3 but lost reactivity when Glu 19 or 20 was changed to Gla. In the order of gamma-carboxylation, MU-3 reacted strongly to DCP containing 0-1 Gla, weakly to 2-4 Gla and not at all to DCP containing more than five Gla. After adsorbing normal prothrombin with barium carbonate, DCP reaction to MU-3 was measured by determining the amount of DCP that was adsorbed by MU-3-coated beads. The proportion of DCP reacting to MU-3 in HCC was 41.0-76.8%, whereas in patients with benign liver diseases, only 0-42.1% reacted to MU-3. These results indicate that DCP variants preferentially synthesized in HCC have less than four Gla, which are restricted to positions 16, 25, 26 and 29, whereas DCP variants in benign liver diseases have more than five Gla.
authors: Tsukuba Research Laboratories, Eisai Co., Ltd., 1-3 Tokodai 5-chome, Ibaraki 300-2635, Japan. t-naraki@hhc.eisai.co.jp
authors: Toru
authors: T
authors: Naraki
authors: Noriatsu
authors: N
authors: Kohno
authors: Hiroyuki
authors: H
authors: Saito
authors: Yoshinori
authors: Y
authors: Fujimoto
authors: Motoyuki
authors: M
authors: Ohhira
authors: Takashi
authors: T
authors: Morita
authors: Yutaka
authors: Y
authors: Kohgo
mesh_terms: D015055:1-Carboxyglutamic Acid / Q000032:analysis*; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D015415:Biomarkers; D001779:Blood Coagulation Factors / Q000276:immunology; D002118:Calcium; D006528:Carcinoma, Hepatocellular; D002413:Cations, Divalent; D002460:Cell Line; D004492:Edetic Acid; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D005164:Factor IX / Q000737:chemistry; D005170:Factor X / Q000737:chemistry; D006801:Humans; D008107:Liver Diseases; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000032:analysis / Q000737:chemistry* / Q000276:immunology; D011516:Prothrombin / Q000032:analysis / Q000737:chemistry* / Q000276:immunology; D051381:Rats; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D001779:Blood Coagulation Factors; D002413:Cations, Divalent; D000939:Epitopes; D011498:Protein Precursors; C008208:acarboxyprothrombin; D015055:1-Carboxyglutamic Acid; D011516:Prothrombin; D005164:Factor IX; D005170:Factor X; D004492:Edetic Acid; D002118:Calcium