RSS-Feed abonnieren
Bitte kopieren Sie die angezeigte URL und fügen sie dann in Ihren RSS-Reader ein.
https://www.thieme-connect.de/rss/thieme/de/10.1055-s-00035024.xml
PDF herunterladen
Thromb Haemost 2001; 86(03): 902-908
DOI: 10.1055/s-0037-1616148
DOI: 10.1055/s-0037-1616148
Review Articles
A Barbourin-albumin Fusion Protein that Is Slowly Cleared In Vivo Retains the Ability to Inhibit Platelet Aggregation In Vitro
* This work was supported by Grant-In-Aid T3730 from the Heart and Stroke Foundation of Ontario.
Weitere Informationen
Publikationsverlauf
Received
30. November 2000
Accepted after resubmission
02. April 2001
Publikationsdatum:
14. Dezember 2017 (online)
Summary
Barbourin is a 73 amino acid venom protein that inhibits platelet aggregation. Recombinant barbourin (BARH6), rabbit serum albumin (RSAH6), and a barbourin-RSA fusion protein (barbourin-linker-albumin; BLAH6) were secreted from Pichia pastoris yeast, and purified by nickel-chelate affinity chromatography via their C-terminal hexahisti-dine (H6) tags. BARH6 and BLAH6 did not differ in their IC50s for inhibition of platelet aggregation using either human platelets stimulated with thrombin or ADP, or rabbit platelets stimulated with ADP. BARH6 and BLAH6 were also effective in inhibiting platelet aggregation in whole blood, and formed complexes with platelet integrin αIIbβ3. The terminal catabolic half-life of BLAH6 approached that of RSAH6[3.4 ± 0.2 versus 4.0 ± 0.1 days (n = 4 ± SD)], but was substantially increased relative to that of BARH6 [0.15 ± 0.03 days (n = 3 ± SD)]. Our results suggest that fusion to albumin slows the clearance of barbourin in vivo, while preserving its ability to inhibit platelet aggregation.
-
References
- 1 George JN. Platelets. Lancet 2000; 355: 1531-9.
- 2 Weiss HJ. Flow-related platelet deposition on subendothelium. Thromb Haemost 1995; 74: 117-22.
- 3 Andrews RK, Lopez JA, Berndt MC. Molecular mechanisms of platelet adhesion and activation. Int J Biochem Cell Biol 1997; 29: 91-105.
- 4 Kulkarni S, Dopheide SM, Yap CL. et al. A revised model of platelet aggregation. J Clin Invest 2000; 105: 783-91.
- 5 Du X, Plow EF, Freelinger III AL. et al. Ligands “activate” integrin αIIbβ3 . Cell 1991; 65: 409-16.
- 6 Plow EF, Byzova T. The biology of glycoprotein IIb-IIIa. Coron Artery Dis 1999; 10: 547-51.
- 7 Coller BS, Scudder LE, Beer J, Gold HK, Folts JD, Cavagnaro J, Jordan R, Wagner C, Iuliucci J, Knight D, Ghrayeb J, Smith C, Weisman HF, Berger H. Monoclonal antibodies to platelet GpIIb/IIIa as antithrombotic agents. Ann NY Acad Sci 1991; 614: 193-213.
- 8 Coller BS, Scudder LE, Beer J. et al. Monoclonal antibodies to platelet GpIIb/IIIa as antithrombotic agents. Ann NY Acad Sci 1991; 614: 193-213.
- 9 Scarborough RM, Kleiman NS, Phillips DR. Platelet glycoprotein IIb/IIIa antagonists. What are the relevant issues concerning their pharmacology and clinical use? Circulation 1999; 100: 437-44.
- 10 Scarborough RM, Rose JW, Hsu MA, Phillips DR, Fried VA, Campbell AM, Nannizzi L, Charo IF. Barbourin. A GPIIb-IIIa-specific integrin antagonist from the venom of Sistrurus m. barbouri . J Biol Chem 1991; 266: 9359-62.
- 11 McLane MA, Marcinkiewicz C, Vijay-Kumar S, Wierzbicka-Patynowski I, Niewiarowski S. Viper venom disintegrins and related molecules. Proc Soc Exp Biol Med 1998; 219: 109-19.
- 12 Scarborough RM. Development of eptifibatide. Am Heart J 1999; 138: 1093-104.
- 13 Scarborough RM, Rose JW, Naughton MA, Phillips DR, Nannizzi L, Arfsten A, Campbell AM, Charo IF. Characterization of the integrin specificities of disintegrins isolated from American pit viper venoms. J Biol Chem 1993; 268: 1058-65.
- 14 Syed S, Schuyler PD, Kulczycky M, Sheffield WP. Potent antithrombin activity and delayed clearance from the circulation characterize recombinant hirudin genetically fused to albumin. Blood 1997; 89: 3243-52.
- 15 Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K. Current Protocols in Molecular Biology. Wiley Interscience; 1989
- 16 Cereghino JL, Cregg JM. Heterologous protein expression in the methylotropic yeast Pichia pastoris. FEMS Microbiol Rev 2000; 24: 45-66.
- 17 Fraker PJ, Speck JC. Protein and cell membrane iodination with a sparingly soluble chloramide – 1,3,4,6-tetrachloro-3,6-diphenylglycoluril. Biochem Biophys Res Comm 1978; 80: 849-52.
- 18 Sheridan D, Carter C, Kelton JG. A diagnostic test for heparin-induced thrombocytopenia. Blood 1986; 67: 27-30.
- 19 Ingerman-Wojenski CM, Silver MJ. A quick method for screening platelet dysfunctions using the whole blood lumi-aggregometer. Thromb Haemost 1984; 51: 154-6.
- 20 Kelton JG, Smith JW, Horsewood P, Humbert JR, Hayward CP, Warkentin TE. Gova/b alloantigen system on human platelets. Blood 1990; 75: 2172-6.
- 21 Marchalonis JJ. An enzymic method for the trace iodination of immunoglobulins and other plasma proteins. Biochem J 1969; 113: 299-305.
- 22 Sheffield WP, Marques JA, Bhakta V, Smith IJ. Modulation of clearance of recombinant serum albumin by either glycosylation or truncation. Thromb Res 2000; 99: 613-21.
- 23 Carson ER, Jones EA. Use of kinetic analysis and mathematical modeling in the study of metabolic pathways in vivo . N Engl J Med 1979; 300: 1016-31.
- 24 Carlson TH, Atencio AC, Simon TB. Behaviour of radioiodinated rabbit antithrombin III. Demonstration of a non-circulating vascular compartment. J Clin Invest 1984; 74: 191-8.
- 25 Harfenist EJ, Packham MA, Mustard JF. Effects of the cell adhesion peptide, Arg-Gly-Asp-Ser, on responses of washed platelets from humans, rabbits, and rats. Blood 1988; 71: 132-6.
- 26 Scarborough RM, Naughton MA, Teng W, Rose JW, Phillips DR, Nannizzi L, Arfsten A, Campbell AM, Charo IF. Design of potent and specific integrin antagonists. Peptide antagonists with high specificity for glycoprotein IIb-IIIa. J Biol Chem 1993; 268: 1066-73.
- 27 Handagama P, Scarborough RM, Shuman MA, Bainton DF. Endocytosis of fibrinogen into megakaryocyty and platelet alpha-granules is mediated by alpha IIb beta 3 (glycoprotein IIb-IIIa). Blood 1993; 82: 135-8.
- 29 Quinn M, Deering A, Stewart M, Cox D, Foley B, Fitzgerald D. Quantifying GP IIb/IIIa receptor binding using two monoclonal antibodies. Circulation 1999; 99: 2231-8.
- 30 Kereiakes DJ, Broderick TM, Roth EM. et al. Time course, magnitude, and consistency of platelet inhibition by abciximab, tirofiban, or eptifibatide in patients with unstable angina pectoris undergoing percutaneous coronary intervention. Am J Cardiol 1999; 84: 391-5.
- 31 Minoux H, Chipot C, Brown D, Maigret B. Structural analysis of the KGD sequence loop of barbourin, an alpha IIb beta3-specific disintegrin. J Comput Aided Mol Des 2000; 14: 317-27.
- 32 Nicolini FA, Lee P, Rios G, Kottke-Marchant K, Topol EJ. Combination of platelet fibrinogen receptor antagonist and direct thrombin inhibitor at low doses markedly improves thrombolysis. Circulation 1994; 89: 1802-9.