Co-expression of tissue inhibitor and matrix metalloproteinase in myocardium
| Autor: | Jeffery Banks, Suresh C. Tyagi, Suresh G. Kumar, Wilbert Fortson |
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| Rok vydání: | 1995 |
| Předmět: |
Male
Molecular Sequence Data Matrix metalloproteinase Chromatography Affinity Rats Sprague-Dawley Interstitial space Affinity chromatography Coumarins Gene expression medicine Animals Zymography Amino Acid Sequence Collagenases RNA Messenger Northern blot Fluorescent Antibody Technique Indirect Molecular Biology In Situ Hybridization Fluorescence Fluorescent Dyes Glycoproteins Chemistry Myocardium Metalloendopeptidases Tissue Inhibitor of Metalloproteinases Tissue inhibitor of metalloproteinase Molecular biology Rats Gene Expression Regulation Gelatinases Chromatography Gel Collagenase Matrix Metalloproteinase 2 Female Matrix Metalloproteinase 1 Cardiology and Cardiovascular Medicine Oligopeptides medicine.drug |
| Zdroj: | Journal of Molecular and Cellular Cardiology. 27:2177-2189 |
| ISSN: | 0022-2828 |
| DOI: | 10.1016/s0022-2828(95)91443-9 |
| Popis: | Matrix metalloproteinases (MMP) are present in the latent form in normal myocardium. To examine the stringent balance between MMP and tissue inhibitor of metalloproteinase (TIMP) and to determine whether MMP are secreted simultaneously and in co-ordination with their inhibotors, we analyzed MMP and TIMP by immunological, isolation br gelpermeation and affinity chromatography, and enzymatic assays in tissues and extracts> Using antibodies to MMP-1 and TIMP-1, we found strong in situ staining of MMP-1 and TIMP-1 in tissues. The staining was uniform in the endo0 and subendomyocardium as well as in the interstitial space. TIMP-1 was present wherever MMP-1 was localized. From the tissue extract, proteins were separated on a gel-filtration column (Sephacryl S-200) and analysed for MMp and TIMP activity by zymography as well as by using succinyl-Gly-Pro-Leu-Gly-Pro-4-amido-7-methyl coumarin (Suc-GPLGP-AMC) as a selective fluorogenic substrate for collagenase. TIMP and MMP were further purified on collagen0Sepharose affinity column. The results indicated that MMP activity was co-eluted with TIMP activity. MMP-1, MMP-2 and TIMP-1 were further analysed by Northern blot for mRNA levels in the heart, skin, lung, liver and kidney. Results suggested co-expression of MMP-1 and TIMP-1 at the transcription level in all tissues. The level of MMP-2 mRNA was specifically higher in the biggest heart tissue, which suggest a role of MMP-2 in the integrity of cardiovascular structure. The study indicate that myocardium as well as other tissue have an endogenous inhibitory system, suggesting that the MMPs activity is co-ordinated by their inhibitors at both the gene and protein levels. Furthermore, MMP and TIMP were co-expressed and were tightly regulated in maintaining the architecture of the interstitial tissue. |
| Databáze: | OpenAIRE |
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