Summary
The objectives of this study were to determine the genetic basis of the electrophoretic differences of human plasma protein C inhibitors (PCI) from 977 individuals. Three discrete antibodies were produced against the PCI purified from human plasma and peptides that corresponded to the N-terminal 15 amino acid residues and the C-terminal 15 residues of human PCI, the chemical structures of which were determined by cDNA sequence analysis. The combined techniques of polyacrylamide gel isoelectric focusing and immunoblotting with these three different antibodies resolved the plasma PCI into several isoprotein bands, with a pH range of 6–7. These PCI isoproteins, however, were not stained by anti-human kallikrein, anti-human protein C or anti-human urokinase antibodies. Therefore, each of the PCI bands, which were detected by immunoblotting with the anti-PCI antibody and the two different anti-peptide antibodies, were derived from free PCI, and not an inactive PCI species. Two common phenotypes, designated PCI 1 and 1–2, were recognized, and family studies showed that they represented homozygosity or heterozygosity for two autosomal codominant alleles, PCI *1 and PCI *2. A population study of plasma samples collected from 977 Japanese individuals indicated that the frequencies of the PCI *1 and PCI *2 alleles were 0.988 and 0.012, respectively.
Similar content being viewed by others
References
España F, Griffin JH (1989) Determination of functional and antigenie protein C inhibitor and its complexes with activated protein C in plasma by ELISA's. Thromb Res 55:671–682
España F, Berrettini M, Griffin JH (1989) Purification and characterization of plasma protein C inhibitor. Thromb Res 55:369–384
Francis RB Jr, Thomas W (1984) Behaviour of protein C inhibitor in intravascular coagulation and liver disease. Thromb Haemost 52:71–74
Heeb MJ, España F, Geiger M, Collen D, Stump DC, Griffin JH (1987) Immunological identity of heparin-dependent plasma and urinary protein C inhibitor and plasminogen activator inhibitor-3. J Biol Chem 262:15813–15816
Heeb MJ, Mosher D, Griffin JH (1989) Activation and complexation of protein C and cleavage and decrease of protein S in plasma of patients with intravascular coagulation. Blood 73:455–461
Kishi K, Yasuda T (1987) Newly characterized genetic polymorphism of uropepsinogen group A (PGA) using both isoelectric focusing and immunoblotting. Hum Genet 75:209–212
Kishi K, Yasuda T, Awazu S, Mizuta K (1989) Genetic polymorphism of human urine deoxyribonuclease I. Hum Genet 81:295–297
Kishi K, Yasuda T, Ikehara Y, Sawazaki K, Sato W, Iida R (1990a) Human serum deoxyribonuclease I (DNase I) polymorphism: pattern similarities among isozymes from serum, urine, kidney, liver, and pancreas. Am J Hum Genet 47:121–126
Kishi K, Yasuda T, Mizuta K (1990b) New genetic markers detected in human urine: DNase I, RNase 1, and 43-kDa glycoprotein. Prog Clin Biol Res 344:891–913
Kuhn LA, Griffin JH, Fisher CL, Greengard JS, Bouma BN, Espana F, Tainer JA (1990) Elucidating the structural chemistry of glycosaminoglycan recognition by protein C inhibitor. Proc Natl Acad Sci USA 87:8506–8510
Laurell M, Stenflo J (1989) Protein C inhibitor from human plasma: characterization of native and cleaved inhibitor and demonstration of inhibitor complexes with plasma kallikrein. Thromb Haemost 62:885–891
Laurell M, Carlson TH, Stenflo J (1988) Monoclonal antibodies against the heparin-dependent protein C inhibitor suitable for inhibitor purification and assay of inhibitor complexes. Thromb Haemost 60:334–339
Marlar RA, Griffin JH (1980) Deficiency of protein C inhibitor in combined factor V/VIII deficiency disease. J Clin Invest 66:1186–1189
Marlar RA, Endres-Brooks J, Miller C (1985) Serial studies of protein C and its plasma inhibitor in patients with disseminated intravascular coagulation. Blood 66:59–63
Meijers JCM, Kanters DHAJ, Vlooswijk RAA, Erp HE van, Hessing M, Bouma BN (1988) Inactivation of human plasma kallikrein and factor XIa by protein C inhibitor. Biochemistry 27:4231–4237
Mizuta K, Yasuda T, Kishi K (1989) Biochemical and genetic studies on GP43, a 43-kD glycoprotein detected immunologically in human urine and serum. Biochem Genet 27:731–743
Morito F, Saito H, Suzuki K, Hashimoto S (1985) Synthesis and secretion of protein C inhibitor by the human hepatoma-derived cell line, Hep G2. Biochim Biophys Acta 844:209–215
Pratt CW, Macik BG, Church FC (1989) Protein C inhibitor: purification and proteinase reactivity. Thromb Res 53:595–602
Stief TW, Radtke K-P, Heimburger N (1987) Inhibition of urokinase by protein C-inhibitor (PCI): evidence for identity of PCI and plasminogen activator inhibitor 3. Biol Chem Hoppe Seyler 368:1427–1433
Suzuki K, Nishioka J, Hashimoto S (1983) Protein C inhibitor: purification from human plasma and characterization. J Biol Chem 258:163–168
Suzuki K, Nishioka J, Kusumoto H, Hashimoto S (1984) Mechanism of inhibition of activated protein C by protein C inhibitor. J Biochem 95:187–195
Suzuki K, Deyashiki Y, Nishioka J, Kurachi K, Akira M, Yamamoto S, Hashimoto S (1987) Characterization of cDNA for human protein C inhibitor: a new member of the plasma serine protease inhibitor superfamily. J Biol Chem 262:611–616
Suzuki K, Deyashiki Y, Nishioka J, Toma K (1989) Protein C inhibitor: structure and function. Thromb Haemost 61:337–342
Yasuda T, Sato W, Mizuta K, Kishi K (1988) Genetic polymorphism of human serum ribonuclease 1 (RNase 1). Am J Hum Genet 42:608–614
Yasuda T, Ikehara Y, Takagi S, Mizuta K, Kishi K (1989a) A hereditary double double-banded variation in the vitamin D-binding protein (GC) system analyzed by immunoblotting: duplication of the 1F and 1A2 genes? Hum Genet 82:89–91
Yasuda T, Mizuta K, Ikehara Y, Kishi K (1989b) Genetic analysis of human deoxyribonucleae I by immunoblotting and the zymogram method following isoelectric focusing. Anal Biochem 183:84–88
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Yasuda, T., Nadano, D., Iida, R. et al. Discovery of a genetic polymorphism of human plasma protein C inhibitor (PCI): genetic survey utilizing isoelectric focusing followed by immunoblotting, immunological and biochemical characterization. Hum Genet 89, 265–269 (1992). https://doi.org/10.1007/BF00220537
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00220537