The epidermal growth factor-like domain from tissue plasminogen activator. Cloning in E.coli, purification and ESR studies of its interaction with human blood platelets

• Autorzy: Pietrucha T. , Stec W.J. , Okruszek A. , Uznanski B. , Koziolkiewicz M. , Wilk A. , Plucienniczak A. , Swiatkowska M. , Cierniewski C.S.
• Języki publikacji: EN

Abstrakty

EN

To examine whether the epidermal growth factor (EGF)-like domain Pr047-Asps7 is involved in the interaction of tissue plasminogen activator (t-PA) with platelets, we have expressed this domain in £. coli. The peptide fragment was produced from a plasmid expression vector as a fusion protein with P*galactosidase Meti-Valm at high yield in eight clones of E. coli. The fusion protein was purified and subjected to mild acid hydrolysis with formic acid, then the peptide Pro47-Asps7, identified by immunoblotting using specific antibodies to t-PA, was isolated by HPLC. After incubation with blood platelets spin labelled with 16-doxylstearic acid or 5-doxylstearic acid, the Pro47-Asps7 peptide fragment reduced fluidity of the membrane lipid bilayer to the same extent as did intact t-PA as indicated by ESR measurements. Our data suggest that the EGF-like domain of t-PA can directly interact with blood platelets and thus it seems to contain those sites of the t-PA molecule that bind the platelet membrane components.

Słowa kluczowe

EN

epidermal growth factor; man; cloning; Escherichia coli; tissue plasminogen; platelet

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1994
• Tom: 41
• Numer: 1
• Opis fizyczny: p.25-34,fig.

Twórcy

autor

Pietrucha T.

• Medical University, ul.Lindley 3, 90-131 Lodz, Poland

autor

Stec W.J.

autor

Okruszek A.

autor

Uznanski B.

autor

Koziolkiewicz M.

autor

Wilk A.

autor

Plucienniczak A.

autor

Swiatkowska M.

autor

Cierniewski C.S.

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