Formation of Stable Complexes Between Urokinase and a Human Urinary Component

Witold Cieplak; David A Hart

doi:10.1055/s-0038-1661232

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1985; 53(01): 036-041
DOI: 10.1055/s-0038-1661232

Original Article

Schattauer GmbH Stuttgart

Formation of Stable Complexes Between Urokinase and a Human Urinary Component

Witold Cieplak

¹The Department of Microbiology, University of Texas Health Science Center at Dallas, Canada

,

David A Hart

²The Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Canada

› Author Affiliations

Abstract

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Summary

Human urine was found to contain multiple species of urokinase (UK)-like plasminogen activator (PA) activity when subjected to concentration and/or dialysis and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymography. Untreated, freshly voided urine contained only M_r = 52,000 and 35,000 UK while dialyzed or undialyzed urine concentrates contained additional PA activity at M_r = 80,000 and 95.000. SDS-PAGE of incubation mixtures of radioiodinated UK (M_r = 52,000 or 35,000) and urine concentrates revealed the presence of radiolabeled complexes with M_rS of 80,000 and 95.000. The bond(s) involved in complex formation was also relatively resistant to heat and reduction. Treatment of radioiodinated UK with the serine proteinase inhibitor, p-nitrophenyl guanidinobenzoate, prior to incubation with dialyzed, concentrated urine prevented formation of the complexes. In addition, the enzymatic activity of the M_r = 80,000 and 95,000 species was unaffected by diisopropyl fluorophosphate. These results indicate that UK forms SDS-stable complexes with a urinary component that has a M_r of approximately 40,000. The results further suggest that these complexes express PA activity when analyzed by SDS- PAGE and zymography.

Key words

Urokinase - Zymography - Complexes - Urinary binding component

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References

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