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Thromb Haemost 1990; 63(02): 246-250
DOI: 10.1055/s-0038-1645203
Original Article
Schattauer GmbH Stuttgart

The Conformation Changes of the Finger Domain of Tissue Type Plasminogen Activator during the Activator-lnhibitor Reaction

Malgorzata Wilczyńska
The Department of Biophysics, Institute of Physiology and Biochemistry, Medical School in Lódź, Lódź, Poland
,
Czeslaw S Cierniewski
The Department of Biophysics, Institute of Physiology and Biochemistry, Medical School in Lódź, Lódź, Poland
› Author Affiliations
Further Information

Publication History

Received 22 November 1989

Accepted after revision 27 November 1989

Publication Date:
02 July 2018 (online)

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Summary

A peptide fragment of tissue plasminogen activator (tPA) corresponding to amino acid residues 4-8 (tPA4−8) was synthesized, coupled to thyroglobulin and injected into rabbits. Antibodies specific to the peptide tPA4−8 were purified immunochemically on the pentapeptide coupled to CNBr-Sepha rose 4B. Anti-tPA4−8 antibodies, reacted with iodinated peptide tPA4−8, showing a relatively high binding affinity (KD = 2.3 × 10−8)

There was no interaction between anti-tPA4 − 8 antibodies and native one- or two-chain tPA. However, reduction of disulfide bonds unmasked the epitope on the heavy chain of tPA which became accessible to anti-tPA4−8 antibodies. Similarly, complex-ing of tPA with Α1arantitrypsin inhibitor resulted in unmasking of the epitope formed by amino acid residues in the positions 4-8.

Presented data suggest that complexing of tPA with inhibitors results in conformational changes occurring in the “finger” domain of tPA molecule and such conformational transition can be detected by antipeptide antibodies. Therefore, anti-tPA4−8 antibodies may be employed as sequence-specific reporter molecules to monitor local conformational changes in tPA molecule.

 

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