Protein S mRNA in Patients with Protein S Deficiency

Summary

A protein S gene polymorphism, detectable by restriction analysis (BstXI) of amplified exonic sequences (exon 15), was studied in seven Italian families with protein S deficiency. In the 17 individuals heterozygous for the polymorphism the study was extended to platelet mRNA through reverse transcription, amplification and densitometric analysis. mRNA produced by the putative defective protein S genes was absent in three families and reduced to a different extent (as expressed by altered allelic ratios) in four families. The allelic ratios helped to distinguish total protein S deficiency (type I) from free protein S deficiency (type IIa) in families with equivocal phenotypes. This study indicates that the study of platelet mRNA, in association with phenotypic analysis based upon protein S assays in plasma, helps to classify patients with protein S deficiency.

• References

• 1 Di Scipio RG, Davie EW. Characterization of protein S, an alpha-carb- oxiglutamic acid containing protein from bovine and human plasma. Biochemistry 1979; 18: 899-904
• 2 Esmon CT. The regulation of natural anticoagulant pathways. Science 1987; 235: 1348-1352
• 3 Fair DS, Marlar SA. Biosynthesis and secretion of factor VII, protein C, protein S, and the protein C inhibitor from a human hepatoma cell line. Blood 1986; 67: 64-70
• 4 Ogura M, Tanabe N, Nishioka J, Suzuki K, Saito H. Biosynthesis and secretion of functional protein S by a human megakaryoblastic cell line. Blood 1987; 70: 301-306
• 5 Walker FJ. Regulation of activated protein C by protein S: the role of phospholipid in factor Va inactivation. J Biol Chem 1981; 256: 11128-11131
• 6 Gardiner JE, McGann MA, Berridge CW, Fulcher CA, Zimmerman TS, Griffin JH. Protein S as a cofactor for activated protein C in plasma and in the inactivation of purified factor VIII:C. Circulation 1984; 70: 205
• 7 Dahlbäck B. Inhibition of protein Ca cofactor function of human and bovine protein S by C4B-binding protein. J Biol Chem 1986; 261: 12022-12027
• 8 Comp PC, Esmon CT. Recurrent venous thromboembolism in patients with a partial deficiency of protein C. N Engl J Med 1984; 311: 1525-1528
• 9 Lundwall A, Dackowski W, Cohen E, Shaffer M, Mahr A, Dahlbäck B, Stenflo J, Wydro R. Isolation and sequence of the cDNA for human protein S, a regulator of blood coagulation. Proc Natl Acad Sci USA 1986; 83: 6716-6720
• 10 Ploos van Amstel HK, van der Zanden AL, Bakker E, Reitsma PH, Bertina RM. Two genes homologous with human protein S cDNA are located on chromosome 3. Thromb Haemost 1987; 58: 982-987
• 11 Schmidel DK, Tatro AV, Phelps LG, Tomczak JA, Long GL. Organization of the human protein S genes. Biochemistry 1990; 29: 7845-7852
• 12 Ploos van Amstel HK, Reitsma PH, van der Logt CP E, Bertina RM. Intron-exon organization of the active human protein S gene PSα and its pseudo-gene PSβ: duplication and silencing during primate evolution. Biochemistry 1990; 29: 7853-7861
• 13 Edenbrandt CM, Lundwall A, Wydro R, Stenflo J. Molecular analysis of the gene for vitamin K dependent protein S and its pseudogene Cloning and partial gene organization. Biochemistry 1990; 29: 7861-7868
• 14 Comp PC. Laboratory evaluation of protein S status. Sem Thromb Haemost 1990; 16: 177-181
• 15 Viganò D’Angelo S, D’Angelo A, Kaufman Jr CE, Sholer C, Esmon CT, Comp PC. Protein S deficiency occurs in the nephrotic syndrome. Ann Intern Med 1987; 107: 42-47
• 16 Malm J, Laurell M, Dahlbäck B. Changes in the plasma levels of vitamin K- dependent protein C and S and of C4b-binding protein during pregnancy and oral contraception. Br J Haematol 1988; 68: 437-443
• 17 Diepstraten CM, Ploos van Amstel HK, Reitsma PH, Bertina RM. A CCA/CCG neutral dimorphism in the codon for Pro 626 of the human protein S gene PSα (PROS1). NucI Ac Res 1991; 19: 5091
• 18 Mannucci PM, Valsecchi C, Krachmalnicoff A, Faioni EM, Tripodi A. Familial dysfunction of protein S. Thromb Haemost 1989; 62: 763-766
• 19 Preda L, Tripodi A, Vaisecchi C, Lombardi A, Finotto E, Mannucci PM. A prothrombin time-based functional assay of protein S. Thromb Res 1990; 60: 19-32
• 20 Tripodi A, Bertina RM, Conard J, Sala N, Pabinger I, Mannucci PM. Multicenter evaluation of three commercial methods for measuring protein S antigen. Thromb Haemost 1992; 68: 149-154
• 21 Faioni EM, Franchi F, Asti D, Sacchi E, Bernardi F, Mannucci PM. Resistance to activated protein C in nine thrombophilic families: interference in a protein S functional assay. Thromb Haemost 1993; 70: 1067-1071
• 22 Marchetti G, Legnani C, Patracchini P, Gemmati D, Ferrati M, Palareti G, Coccheri S, Bernardi F. Study of a protein S gene polymorphism at DNA and mRNA level in a family with symptomatic protein S deficiency. Br J Haematol 1993; 85: 173-175
• 23 Ploos van Amstel HK, Diepstraten CM, Reitsma PH, Bertina RM. Analysis of platelet protein S mRNA suggests silent alleles as a frequent cause of hereditary protein S deficiency type I. Thromb Haemost 1991; 65: 808