Identification of prolyl oligopeptidase as a cyclosporine-sensitive protease by screening of mouse liver extracts
| Autor: | Miroslav Malesevic, Alexander Rentzsch, Mike Schutkowski, Gunter Fischer, Günther Jahreis, Viktoria Kahlert, Bernhard Schlott |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
medicine.medical_treatment
Proteolysis Clinical Biochemistry Oligopeptidase Peptide Biochemistry Cyclophilins Mice Immune system Peptide Library medicine Animals Humans Peptide library Receptor Molecular Biology Cyclophilin chemistry.chemical_classification Protease medicine.diagnostic_test Chemistry Serine Endopeptidases Mice Inbred C57BL Cyclosporine Liver Extracts Prolyl Oligopeptidases Immunosuppressive Agents |
| Zdroj: | bchm. 394:1057-1067 |
| ISSN: | 1437-4315 1431-6730 |
| DOI: | 10.1515/hsz-2013-0125 |
| Popis: | Cyclosporine A (CsA) is a cyclic undecapeptide well known for its ability to prevent rejection episodes after organ transplantation via gain-of-function. Therefore, biomedical studies on CsA have been focused on both immunosuppressive properties and binding to the biocatalytically-active immune receptors, the cyclophilins. Much less attention has been spent on effects of cyclosporines on the biological function of other proteins. We used a 9-mer fluorescence-quenched peptide library with defined sequences to identify cyclosporine-sensitive proteolysis in mouse liver extracts. A highly soluble [d-Ser]8-CsA derivative was utilized to avoid drug precipitation at extended incubation times. Analysis of the time courses of proteolysis revealed 15 out of 360 peptide sequences where proteolysis exhibited marked sensitivity to the cyclosporine derivative. As a first example, a collagen-derived substrate was selected from those hits to identify the targeted proteolytic pathway. After purification from mouse liver extracts, prolyl oligopeptidase (EC 3.4.21.26) could be identified as a protease sensitive to submicromolar concentrations of cyclosporines. Surprisingly, in a series of cyclosporine derivatives an inverse relationship was found between the inhibition of prolyl oligopeptidase and inhibition of cyclophilin A. |
| Databáze: | OpenAIRE |
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