Fluorescence Polarization Assay and SDS-PAGE Confirms Matrilysin Degrades Fibronectin and Collagen IV whereas Gelatinase: A Degrades Collagen IV but not Fibronectin
| العنوان: | Fluorescence Polarization Assay and SDS-PAGE Confirms Matrilysin Degrades Fibronectin and Collagen IV whereas Gelatinase: A Degrades Collagen IV but not Fibronectin |
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| المؤلفون: | Donna E. Haynes-Johnson, Lekha Patel, Judy A. Lenhart, Stephen S. Palmer, Patricia Kraft, Zivin Robert |
| المصدر: | Connective Tissue Research. 42:149-163 |
| بيانات النشر: | Informa UK Limited, 2001. |
| سنة النشر: | 2001 |
| مصطلحات موضوعية: | Collagen Type IV, medicine.medical_treatment, Proteolysis, Gelatinase A, Fluorescence Polarization, Peptide, Biochemistry, Substrate Specificity, Rheumatology, Cricetinae, medicine, Animals, Humans, Orthopedics and Sports Medicine, Matrilysin, Molecular Biology, chemistry.chemical_classification, Protease, medicine.diagnostic_test, biology, Cell Biology, Molecular biology, In vitro, Fibronectins, Fibronectin, Animals, Newborn, chemistry, Matrix Metalloproteinase 7, biology.protein, Matrix Metalloproteinase 2, Electrophoresis, Polyacrylamide Gel, Fluorescence anisotropy |
| الوصف: | Matrilysin and gelatinase A are hypothesized to have significant roles in uterine and ovarian function. However, proteolytic activity assays for these enzymes are limited. We describe the development of simple and rapid assays for the proteolysis of fluorescein-labeled full-length substrates, collagen IV (Col-IV) and fibronectin (FN), and demonstrate the selectivity of matrilysin (MMP-7) compared to gelatinase A (MMP-2) for fibronectin. Changes in fluorescence intensity (FIU) and fluorescence polarization (mP) resulting from the protease activity of matrilysin and gelatinase A were measured. These studies show that the fluorescently labeled substrates, Col-IV and FN, are as reliable and amenable to rapid in vitro assay as peptide substrates. In addition, they are easier to use than previously described, non-fluorescent methods. The results demonstrate that assays using full-length, biological matrix proteins are more sensitive indicators of MMP-specific substrate activity than peptide based assays. |
| تدمد: | 1607-8438 0300-8207 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e7d7bc6a7efd341309c3f5f5054f653aTest https://doi.org/10.3109/03008200109014256Test |
| رقم الانضمام: | edsair.doi.dedup.....e7d7bc6a7efd341309c3f5f5054f653a |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 16078438 03008207 |
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