Die molekularbiologischen Eigenschaften des Gewebetyp-Plasminogenaktivators und seiner Varianten

P. Tanswell; J. Krause

doi:10.1055/s-0038-1655260

Hämostaseologie, Inhaltsverzeichnis

Hamostaseologie 1989; 09(03): 114-121
DOI: 10.1055/s-0038-1655260

Originalarbeiten

Schattauer GmbH

Die molekularbiologischen Eigenschaften des Gewebetyp-Plasminogenaktivators und seiner Varianten

Authors

P. Tanswell

¹Abteilung Biochemie, Dr. Karl Thomae GmbH, Biberach an der Riß
J. Krause

¹Abteilung Biochemie, Dr. Karl Thomae GmbH, Biberach an der Riß

Abstract

Zusammenfassung

Der Gewebetyp-Plasminogenaktivator (t-PA) ist ein Schlüsselenzym der physiologischen Fibrinolyse und steht heute auch als rekombinantes Protein für therapeutische Zwecke zur Verfügung. Biotechnisch hergestelltes t-PA (Actilyse®) verzeichnet bereits beachtliche klinische Erfolge bei der Behandlung von thromboembolischen Erkrankungen, insbesondere dem akuten Herzinfarkt. t-PA aktiviert Plasminogen zu Plasmin, welches das Fibringerüst eines Thrombus auflösen kann. Diese Aktivierung erfolgt selektiv auf der Thrombusoberfläche und verursacht nur geringe systemische Nebenwirkungen. Die Halbwertszeit von t-PA in vivo ist aufgrund einer raschen hepatischen Elimination sehr kurz (3-5 min).

t-PA ist ein Glykoprotein mit Serinproteaseaktivität, dessen Polypeptidkette aus 527 Aminosäuren besteht und über insgesamt 17 Disulfidbrükken verknüpft ist. Mit dem Ziel, Fibrinspezifität und katalytische Aktivität zu erhöhen oder die Eliminationsrate zu verringern, haben zahlreiche Forschergruppen in jüngster Zeit Modifikationen der molekularen Struktur von t-PA durchgeführt. Durch diese Arbeiten wurden zwar interessante Beziehungen zwischen Struktur und Funktion des t-PA-Moleküls aufgedeckt, jedoch konnte bisher keine klinisch relevante Variante konstruiert werden, die dem natürlich vorkommenden Protein in der Summe seiner Eigenschaften überlegen ist.

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Referenzen

LITERATUR
1 Collen D. On the regulation and control of fibrinolysis. Thromb Haemostas 1980; 43: 77-89.
2 Emeis J J, Brommer E J P, Kluft C, Brakman R. Progress in Fibrinolysis. In: Recent Advances in Blood Coagulation 4. Poller L. (Ed). Edinburgh: Churchill Livingstone; 1985: 11-34.
3 Kluft C. Plasminogen proactivators and activators in human plasma; Factor XII-dependent fibrinolysis. In: Protides of the Biological Fluids. Vol XXVIII. Peters H. (Ed). Oxford: Pergamon Press; 1980: 375-8.
4 Blasi F. Surface receptors for urokinase plasminogen activator. Fibrinolysis 1988; 02: 73-84.
5 Layer G T, Cederholm-Williams S A, Gaffney P J, Houlbrook S, Mahmoud M, Pattison M, Burnand K G. Urokinase the enzyme responsible for invasion and metastasis in human breast carcinoma?. Fibrinolysis 1987; 01: 237-40.
6 Aoki N, Harpel P C. Inhibitors of the fibrinolytic enzyme system. Sem Thromb Haemostas 1984; 10: 24-41.
7 Sprengers E D, Kluft C. Plasminogen activator inhibitors. Blood 1987; 69: 381-7.
8 DeWood M A, Spores J, Notske R. et al. Prevalence of total coronary occlusion during the early hours of transmural myocardial infarction. N Engl J Med 1980; 303: 897-901.
9 Sobel B E, Collen D, Grossbard E B. (Eds). Tissue Plasminogen Activator in Thrombolytic Therapy. New York: Marcel Dekker; 1987
10 Verstraete M, Bory M, Collen D, Erbel R. et al. Randomised trial of intravenous recombinant tissue-type plasminogen activator versus intravenous strepokinase in acute myocardial infarction. Lancet 1985; 8433: 842-7.
11 Autoren Verschiedene. Neothrombolytika: rt-PA (Actilyse®). Hämostaseologie 1988; 08: 199-239.
12 Seifried E, Lechner K, Heimpel H. (Eds). Neuere Entwicklungen in der fibrinolytischen Therapie. Klin Wochenschr. 1988. 66 (Suppl XII): 1-149.
13 Hoylaerts M, Rijken D C, Lijnen H R, Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator: role of fibrin. J Biol Chem 1982; 257: 2912-9.
14 Krause J. Catabolism of tissue type plasminogen activator (t-PA), its variants, mutants and hybrids. Fibrinolysis 1988; 02: 133-42.
15 Pennica D, Holmes W E, Kohr W J, Harkins R N. et al. Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli. Nature 1983; 301: 214-21.
16 Ny T, Elgh F, Lund B. The structure of the human tissue-type plasminogen activator gene : correlation of intron and exon structures to functional and structural domains. Proc Natl Acad Sei USA 1984; 81: 5355-9.
17 Friezner S JDegen, Rajput B, Reich E. The human tissue plasminogen activator gene. J Biol Chem 1986; 261: 6972-85.
18 Pohl G, Källstrom M, Bergsdorf N, Wallen P, Jörnvall H. Tissue plasminogen activator: peptide analyses confirm an indirectly derived amino acid sequence, identify the active site serine residue, establish glycosylation sites, and identify variant differences. Biochemistry 1984; 23: 3701-7.
19 Rijken D C, Hoylaerts M, Collen D. Fibrinolytic properties of one-chain and twochain human extrinsic (tissue-type) plasminogen activator. J Biol Chem 1982; 257: 2910-5.
20 Spellman M W, Ferguson C K, O’Connor J V. Characterization of N-linked oligosaccharides from recombinant tissue-tpye plasminogen activator. XIII International Carbohydrate Symposium 1986, Cornell University. New York: Ithaca, Abstr; B122.
21 Pohl G, Kenne L, Nilsson B, Einarsson M. Isolation and characterization of three different carbohydrate side chains from melanoma tissue plasminogen activator. Eur J Biochem 1987; 170: 69-75.
22 Pathy L. Evolution of the proteases of blood coagulation and fibrinolysis by assembly from modules. Cell 1985; 41: 657-63.
23 Banyai L, Varadi A, Patthy L. Common evolutionary origin of the fibrin binding structures of fibronectin and tissue-type plasminogen activator. FEBS Lett 1983; 163: 37-41.
24 Savage C R, Inagami T, Cohen S. The primary structure of epidermal growth factor. J Biol Chem 1972; 247: 7612-21.
25 Sottrup-Jensen L, Claeys H, Zajdel M, Petersen T E, Magnusson S. The primary structure of human plasminogen. In: Progress in Chemical Fibrinolysis and Thrombolysis. Davidson J F, Rowan R M, Samama M M, Desuoyers P C. (Eds), Vol 3, 191-209 New York: Raven Press; 1978
26 Panekoek H, de Vries C, van Zonneveld A J. Mutants of tissue-type plasminogen activator (t-PA): structural aspects and functional properties. Fibrinolysis 1988; 02: 123-32.
27 Bachmann F. Struktur, Funktion und Anwendung von Fibrinolyse-fördernden und -inhibierenden Faktoren. Arzneim-Forsch/ Drug Res 1988; 38 (I): 474-8.
28 Hotchkiss A, Refino C J, Leonard C K, O’Connor J V, Cröwley C, McCabe J, Tata K, Nakamura G, Powers D, Levinson A, Mohler M, Spellmann M W. The influence of carbohydrate structure on the clearance of recombinant tissue-type plasminogen activator. Thromb Haemostas 1988; 60: 255-61.
29 Klausner A. Second generation t-PA race heats up. Biotechnology 1987; 05: 869-70.
30 Smedsrod B, Einarsson M, Pertoft H. Tissue plasminogen activator is endocytosed by mannose and galactose receptors of rat liver cells. Thromb Haemostas 1988; 59: 480-4.
31 Lucore C L, Fry E T A, Nachowiak D A, Sobel B E. Biochemical determinants of clearance of tissue-type plasminogen activator from the circulation. Circulation 1988; 77: 906-14.
32 Kuiper J, Otter M, Rijken D C, van Berkel T J C. Characterization of the interaction in vivo of tissue-type plasminogen activator with liver cells. J Biol Chem 1988; 263: 18220-4.
33 Tate K M, Higgins D L, Holmes W E, Winkler M E, Heynecker H L, Vehar G A. Functional role of proteolytic cleavage at arginine-275 of human tissue plasminogen activator as assessed by site-directed mutagenesis. Biochemistry 1987; 26: 338-43.
34 Petersen L C, Johannessen M, Foster D, Kumar A, Mulvihill E. The effect of polymerised fibrin on the catalytic activities of one-chain tissue-type plasminogen activator as revealed by an analogue resistant to plasmin cleavage. Biochim Biophys Acta 1988; 952: 245-54.
35 Astedt B, Bladh B, Christensen U, Lecander I. Different inhibition of one and two chain tissue plasminogen activator by a placental inhibitor studied with two tripeptide-p-nitro-anilide substrates. Scand J Lab Invest 1985; 45: 429-34.
36 Haigwood N, Paques E-P, Mullenbach G, Moore G, Des Jardin L, Tabrizi A. Improvement of t-PA properties by means of site directed mutagenesis. Thromb Haemostas 1987; 58 Abstr 1042, 286.
37 Häggroth L, Mattsson C, Felding P, Nilsson I M. Plasminogen activator inhibitors in plasma and platelets from patients with recurrent venous thrombosis and pregnant women. Thromb Res 1986; 42: 585-94.
38 Rânby M, Bergsdorf N, Nilsson T. Enzymatic properties of the oneand two-chain form of tissue plasminogen activator. Thromb Res 1982; 27: 175-83.
39 Browne M J, Carey J E, Chapman C G, Tyrrell A W R, Entwisle C, Lawrence B R, Dodd I, Esmail A, Robinson J H A. Tissuetype plasminogen activator mutant with prolonged clearance in vivo. J Biol Chem 1988; 263: 1599-602.
40 Dodd I, Nunn B, Robinson J H. Isolation, identification and pharmacokinetic properties of human tissue-type plasminogen activator species: possible localisation of a clearance recognition site. Thromb Haemostas 1988; 59: 523-8.
41 Fu K P, Lee S, Hum W T, Kalyan N, Rappaport R, Hetzel N, Hung P P. Disposition of a novel recombinant tissue plasminogen activator delta 2-89 t-PA, in mice. Thromb Res 1988; 50: 33-41.
42 Kalyan N K, Lee S G, Wilhelm J, Fu K P, Hum W-T, Rappaport R, Hartzell R W, Urbano C, Hung P P. Structure-function analysis with tissue-type plasminogen activator. J Biol Chem 1988; 263: 3971-8.
43 Larsen G R, Hensen K, Blue Y, Horgan P. The finger domain: a primary determinant in the clearance of t-PA in rat. Fibrinolysis 1988; 02 (Suppl. 01) 29.
44 Johannessen M, Diness V, Pingel K, Petersen L C, Rao D, Insley M, Mulvihill E. Recombinant t-PA deletion analogues: activity, fibrin binding and clearance studies. Fibrinolysis 1988; 02 (Suppl. 01) 30.
45 Collen D, Stassen J-M, Larsen G. Pharmacokinetics and thrombolytic properties of deletion mutants of human tissue-type plasminogen activator in rabbits. Blood 1988; 71: 216-9.
46 Cambier P, Van de Werf F, Larsen G R, Collen D. Pharmacokinetics and thrombolytic properties of a nonglycosylated mutant of human tissue-type plasminogen activator, lacking the finger and growth factor domains, in dogs with copper coil-induced coronary artery thrombosis. J Cardiovasc Pharm 1988; 11: 468-72.
47 van Zonneveld A-J, Yeerman H, Pannekoek H. Autonomous functions of structural domains on human-tissue-type plasminogen activator. Proc Natl Acad Sci 1986; 83: 4670-4.
48 Kagitani H, Tagawa M, Hatanaka K, Ikari T, Saito A, Bando H, Okada K, Matsuo O. Expression in E. coli of finger-domain lakking tissue-type plasminogen activator with high fibrin affinity. FEBS 1985; 189: 145-9.
49 Larsen G R, Henson K, Blue Y. Variants of human tissue-type plasminogen activator. J Biol Chem 1988; 263: 1023-9.
50 Ehrlich H J, Bang N U, Little S P, Jaskunas S R, Weigel B J, Mattier L E, Harms C S. Biological properties of a kringleless tissue plasminogen activator (t-PA) mutant. Fibrinolysis 1987; 01: 75-81.
51 Bergum P W, Erickson L A. Neutralization by plasminogen activator inhibitor-1 of mutants of tissue plasminogen activator. Enzyme 1988; 40: 122-9.
52 Tanswell P, Schlüter M, Krause J. Pharmacokinetics and isolated liver perfusion of carbohydrate-modified recombinant tissuetype plasminogen activator. Fibrinolysis. 1989 (in press).
53 Beebe D P, Aronson D L. Turnover of t-PA in rabbits: influence of carbohydrate moieties. Thromb Res 1988; 51: 11-22.
54 Little S P, Bang U, Harms C S, Marks C A, Mattier L E. Functional properties of carbohydrate-depleted tissue plasminogen activator. Biochem 1984; 23: 6191-5.
55 Sambrook J, Hanahan D, Rodgers L, Gething M-J. Expression of human tissue-type plasminogen activator from lytic viral vectors and in established cell lines. Mol Biol Med 1986; 03: 459-81.
56 Nelles L, Lijnen H R, Collen D, Holmes W E. Characterization of a fusion protein consisting of amino acids 1 to 263 of tissuetype plasminogen activator and amino acids 144 to 411 of urokinase-type plasminogen activator. J Biol Chem 1987; 262: 10855-62.
57 Piérard L, Jacobs P, Gheysen D, Hoylaerts M, André B, Topisirovic L, Cravador A, de Foresta F, Herzog A, Collen D, De Wilde M, Bollen A. Mutant and chimeric recombinant plasminogen activators. J Biol Chem 1987; 262: 11771-8.
58 Gheysen D, Lijnen H R, Piérard L, de Foresta F, Demarsin E, Jacobs P, De Wilde M, Bollen A, Collen D. Characterization of a recombinant fusion protein of the finger domain of tissue-type plasminogen activator with a truncated single chain urokinase-type plasminogen activator. J Biol Chem 1987; 262: 11779-84.
59 Lee S G, Kalyan N, Wilhelm J, Hum W-T, Rappaport R, Cheng S-M, Dheer S, Urbano C, Hartzell R W, Ronchetti-Blume M, Levner M, Hung P P. Construction and expression of hybrid plasminogen activators prepared from tissue-type plasminogen activator and urokinase-type plasminogen activator genes. J Biol Chem 1988; 263: 2917-24.
60 de Vries C, Veerman H, Blasi F, Pannekoek H. Artificial exon shuffling between tissue-type plasminogen activator (t-PA) and urokinase (u-PA): a comparative study on the fibrinolytic properties of t-PA/u-PA hybrid proteins. Biochem 1988; 27: 2565-72.
61 Fu K P, Lee S, Hetzel N, Fenichel R, Hum H W, Speth J, Kalyan N, Hung P P. Clearance of a novel recombinant tissue plasminogen activator in rabbits. Thromb Res 1988; 50: 679-85.
62 Schnee J M, Runge M S, Matsueda G R, Hudson N W, Seidman J G, Haber E, Quertermous T. Construction and expression of a recombinant antibody-targeted plasminogen activator. Proc Natl Acad Sei 1987; 84: 6904-8.
63 Lee P P, Wohl R C, Boreisha I G, Robbins K C. Kinetic analysis of covalent hybrid plasminogen activators: effect of CNBrdegraded fibrinogen on kinetic parameters of Glui-plasminogen activation. Biochem 1988; 27: 7506-13.
64 Robinson J H, Dodd I, Esmail A, Ferres H, Nunn B. Slow clearance of acylated, hybrid thrombolytic enzymes. Thromb Haemostas 1988; 59: 421-5.
65 Runge M A, Bode C, Matsueda G R, Haber E. Conjugation to an antifibrin monoclonal antibody enhances the fibrinolytic potency of tissue plasminogen activator in vitro. Biochem 1988; 27: 1153-7.
66 Runge M A, Bode C, Matsueda G R, Haber E. Antibody-enhanced thrombolysis: targeting of tissue plasminogen activator in vivo. Proc Natl Acad Sci 1987; 84: 7659-62.
67 Kalindjian S B, Smith A G. Reagents for reversible coupling of proteins to the active centres of trypsin-like serine proteinases. Biochem 1987; 248: 409-13.
68 Cassels R, Fears R, Smith A G. The interaction of streptokinase-plasminogen activator complex, tissue-type plasminogen activator, urokinase and their acylated derivatives with fibrin and cyanogen bromide digest of fibrinogen. Biochem 1987; 247: 395-400.
69 Stürzebecher J, Richter M, Markwardt F. Stable acyl derivatives of tissue-type plasminogen activator. Thromb Res 1987; 47: 699-703.
70 Garman A J, Kalindjian S B. The preparation and properties of novel reversible polymer-protein conjugates. FEB 1987; 223: 361-5.
71 Berger Jr H, Pizzo S V. Preparation of polyethylene glycol-tissue plasminogen activator adducts that retain functional activity: characteristics and behavior in three animal species. Blood 1988; 71: 1641-7.
72 MacDonald M E, van Zonnefeld A-J, Pannekoek H. Functional analysis of the human tissue-type plasminogen activator protein: the light chain. Gene 1986; 42: 59-67.