Download PDF
Thromb Haemost 1995; 74(06): 1491-1496
DOI: 10.1055/s-0038-1649971
Original Articles
Fibrinolysis
Schattauer GmbH Stuttgart

Identification and Genetic Analysis of a Common Molecular Variant of Histidine-rich Glycoprotein with a Difference of 2KD in Apparent Molecular Weight

B C Hennis
The Gaublus Laboratory TNO-PG, Leiden, The Netherlands
,
P A van Boheemen
The Gaublus Laboratory TNO-PG, Leiden, The Netherlands
,
S Wakabayashi
1   The Department of Life Science, Himeji Institute of Technology, Kamigorl, Japan
,
T Koide
1   The Department of Life Science, Himeji Institute of Technology, Kamigorl, Japan
,
J J M L Hoffmann
2   The Haemostasis Division, Department of Clinical Laboratories, Catharina Hospital, Eindhoven, Leiden, The Netherlands
,
P Klevit
The Gaublus Laboratory TNO-PG, Leiden, The Netherlands
,
G Dooijewaard
The Gaublus Laboratory TNO-PG, Leiden, The Netherlands
,
J G Jansen
3   The MGC-Department of Radiation Genetics and Chemical Mutagenesis, State University Leiden, Leiden, The Netherlands
,
C Kluft
The Gaublus Laboratory TNO-PG, Leiden, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 21 March 1995

Accepted after resubmission 11 August 1995

Publication Date:
27 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

Two forms of histidine-rich glycoprotein (HRG) were detected on SDS-PAGE by silver staining and immunoblotting after isolation of the protein from pooled plasma using immuno-affinity chromatography followed by chromatography with heparin-Sepharose. Both forms were single-chain molecules and the apparent molecular weights of form 1 and form 2 were 77 kD and 75 kD respectively. Mendelian inheritance of both HRG forms was observed in four families with 24 informative meioses, strongly suggesting that the two forms are encoded by different alleles. The frequency of form 1 and form 2 in a group of 36 individuals was 0.35 and 0.65 respectively.

The difference between the two molecular variants was studied by direct sequence analysis of amplified exons of the HRG gene from 6 individuals who were homozygous either for form 1 or form 2. Five amino acid polymorphisms in three different exons were observed: Ile/Thr in exon 4; Pro/Ser in exon 5; His/Arg, Arg/Cys and Asn/Ile in exon 7. Analysis of these polymorphisms in 20 volunteers showed that only the Pro/Ser polymorphism at position 186 in exon 5 was coupled to the form of the HRG protein. Ser was found in form 1 and Pro in form 2. The presence of Ser at position 186 introduces a consensus sequence for a N-glycosylation site (Asn-X-Ser/Thr). By removing N-linked sugars with N-glycanase, it could be demonstrated that the difference between the two forms of HRG is caused by an extra carbohydrate group at Asn 184 in form 1.

 

  • References

  • 1 Haupt H, Heimburger N. Human serum proteins with a high affinity to carboxymethylcellulose, I: Isolation of lysozyme, Clq and two hitherto unknown α-globulins. Hoppe Seylers Z Physiol Chem 1972; 353: 1125-1132
    CrossrefPubMedSearch in Google Scholar
  • 2 Heimburger N, Haupt H, Kranz T, Baudner S. Human serum proteins with a high affinity to carboxymethylcellulose, II: Physico-chemical and immunological characterization of a histidine-rich 3,8S-2-glycoprotein (CM-protein I). Hoppe Seylers Z Physiol Chem 1972; 353: 1133-1140
    CrossrefPubMedSearch in Google Scholar
  • 3 Leung LL K, Harpel PC, Nachman RL, Rabellino EM. Histidine-rich glycoprotein is present in human platelets and is released following thrombin stimulation. Blood 1983; 62: 1016-1021
    PubMedSearch in Google Scholar
  • 4 Hennis BC, De Maat MP M, Quax PH A, Le Clercq EJ, Kuiper J, Kluft C. Evaluation of sites of synthesis of the histidine-rich glycoprotein. Thromb Haemost 1991; 65: 660a abstr
    PubMedSearch in Google Scholar
  • 5 Lijnen HR, Hoylaerts M, Collen D. Heparin binding properties of human histidine-rich glycoprotein. Mechanism and role in the neutralization of heparin in plasma J Biol Chem 1983; 258: 3803-3808
    PubMedSearch in Google Scholar
  • 6 Leung LL K. Interaction of histidine-rich glycoprotein with fibrinogen and fibrin. J Clin Invest 1986; 77: 1305-1311
    CrossrefPubMedSearch in Google Scholar
  • 7 Lijnen HR, Hoylaerts M, Collen D. Isolation and characterization of a human plasma protein with affinity for the lysine binding sites in plasminogen. Role in the regulation of fibrinolysis and identification as histidine-rich glycoprotein J Biol Chem 1980; 255: 10214-10222
    PubMedSearch in Google Scholar
  • 8 Lerch PG, Nydegger UE, Kuyas C, Haeberli A. Histidine-rich glycoprotein binding to activated platelets. Br J Haematol 1988; 70: 219-224
    CrossrefPubMedSearch in Google Scholar
  • 9 Koide T. The primary structure of human histidine-rich glycoprotein and its functions as a modulator of coagulation and fibrinolysis. In: Fibrinolysis: Current Prospects. Gaffney PJ. ed London: John Libbey & Co; 1988. p 55-63
    Search in Google Scholar
  • 10 Morgan WT. Human serum histidine-rich glycoprotein. I. Interactions with heme, metal ions and organic ligands Biochim Biophys Acta 1978; 535: 319-333
    CrossrefPubMedSearch in Google Scholar
  • 11 Shatsky M, Saigo K, Burdach S, Leung LL K, Levitt LJ. Histidine-rich glycoprotein blocks T cell rosette formation and modulates T cell activation and immunoregulation. J Biol Chem 1989; 264: 8254-8259
    PubMedSearch in Google Scholar
  • 12 Muldbjerg M, Shousboe I, Halkier T. Purification of bovine histidine-rich glycoprotein. Thromb Res 1992; 65: 815-819
    CrossrefPubMedSearch in Google Scholar
  • 13 Morgan WT. Interactions of the histidine-rich glycoprotein of serum with metals. Biochemistry 1981; 20: 1054-1061
    CrossrefPubMedSearch in Google Scholar
  • 14 Shimada K, Kawamoto A, Matsubayashi K, Ozawa T. Histidine-rich glycoprotein does not interfere with interactions between antithrombine III and heparin-like compounds on vascular endothelial cells. Blood 1989; 73: 191-193
    PubMedSearch in Google Scholar
  • 15 Sia DY, Rylatt DB, Parish CR. Anti-self receptors. V. Properties of a mouse serum factor that blocks autorosetting receptors on lymphocytes Immunology 1982; 45: 207-216
    PubMedSearch in Google Scholar
  • 16 Koide T, Foster D, Yoshitake S, Davie EW. Amino acid sequence of human histidine-rich glycoprotein derived from the nucleotide sequence of its cDNA. Biochemistry 1986; 25: 2220-2225
    CrossrefPubMedSearch in Google Scholar
  • 17 Yip TT, Hutchens TW. Metal ion affinity adsorption of a Zn(II)-transport protein present in maternal plasma during lactation: structural characterization and identification as histidine-rich glycoprotein. Protein Expression and Purification 1991; 2: 355-362
    CrossrefPubMedSearch in Google Scholar
  • 18 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedSearch in Google Scholar
  • 19 Koide T, Odani S, Ono T. Human histidine-rich glycoprotein: Simultaneous purification with antithrombin III and characterization of its gross structure. J Biochem 1985; 98: 1191-1200
    CrossrefPubMedSearch in Google Scholar
  • 20 Leung LL K, Saigo K, Grant D. Heparin binds to human monocytes and modulates their procoagulant activities and secretory phenotypes. Effect of histidine-rich glycoprotein Blood 1989; 73: 177-184
    PubMedSearch in Google Scholar
  • 21 Schägger H, von Jagow G. Tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis for the separation of proteins in the range 1 to 100 kDa. Anal Biochem 1987; 166: 368-379
    CrossrefPubMedSearch in Google Scholar
  • 22 Wray W, Boulikas T, Wray VP, Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem 1981; 118: 197-203
    CrossrefPubMedSearch in Google Scholar
  • 23 Wijmenga C, Frants RR, Brouwer OF, Van der Klift HM, Meera Khan P, Padberg GW. Facioscapulohumeral muscular gene in Dutch families is not linked to markers for familial adenomatous polyposis on the long arm of chromosome 5. J Neur Sci 1990; 95: 225-229
    CrossrefPubMedSearch in Google Scholar
  • 24 Menichini P, Vrieling H, van Zeeland AA. Strand-specific mutation spectra in repair-proficient and repair deficient hamster cells. Mutation Res 1991; 251: 143-155
    CrossrefPubMedSearch in Google Scholar
  • 25 Russ AP, Maerz W, Ruzicka V, Stein U, Groβ W. Rapid detection of the hypertension-associated Met235→Thr allele of the human angiotensinogen gene. Hum Mol Gen 1993; 2: 609-610
    CrossrefPubMedSearch in Google Scholar
  • 26 Peake I, Winship P. What is a polymorphism. Blood 1991; 78: 251
    PubMedSearch in Google Scholar
  • 27 Sørensen CB, Krogh-Pedersen H, Petersen TE. Determination of the disulphide bridge arrangement of bovine histidine-rich glycoprotein. FEBS Lett 1993; 328: 285-290
    CrossrefPubMedSearch in Google Scholar