Complexes of tissue kallikrein with protein C inhibitor in human semen and urine
| Autor: | Francisco España, Jaime Sánchez-Cuenca, Karlheinz Witzgall, Amparo Estellés, Juan Gilabert, Edwin Fink |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Adult
Male Protein C inhibitor Tissue kallikrein Semen Enzyme-Linked Immunosorbent Assay Urine Biochemistry medicine Humans cardiovascular diseases Protein C Inhibitor Chromatography biology Heparin Middle Aged Molecular biology Urokinase-Type Plasminogen Activator In vitro surgical procedures operative Conventional PCI biology.protein Kallikreins Antibody therapeutics Tissue Kallikreins medicine.drug |
| Zdroj: | European journal of biochemistry. 234(2) |
| ISSN: | 0014-2956 |
| Popis: | An ELISA was developed for quantifying the complex between tissue kallikrein (tKK) and protein C inhibitor (PCI) (tKK:PCI) in seminal plasma and urine. The ELISA used purified tKK:PCI complex as a standard and was specific for this complex with a detection limit of about 1.1 pM. Purified tKK:PCI complex was obtained from human urine and was 95% homogeneous as judged by SDS/PAGE. The 90-kDa band corresponding to the purified tKK:PCI complex reacted with anti-tKK and anti-PCI antibodies as judged by immunoblotting. Seminal plasma collected in the absence of extrinsic inhibitors contained 1.8 +/- 0.6 nM tKK:PCI complex and 4.7 +/- 2.8 nM immunoreactive tKK (mean +/- SD, n = 10), which indicates that about 28% of the total tKK immunoreactivity is forming complexes with PCI. When semen was collected in the presence of tKK inhibitors it had only about 6% of the tKK complexed to PCI. In vitro studies showed that the tKK:PCI complex formation in semen was accomplished in about 1 h and that heparin stimulated both the rate and the extent of complexation of tKK with PCI. Native urine showed low levels of tKK:PCI complex, but after dialysis urine had 0.17 +/- 0.05 nM complex. Formation of tKK:PCI complex in urine and semen was also demonstrated by immunoblotting. These results suggest that PCI is a physiological inhibitor of tKK and provide additional evidence of the involvement of PCI in human reproduction. |
| Databáze: | OpenAIRE |
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