Isolation of Latent Matrix Metallo Proteinase-1 (Latent Collagenase-1) from Serum and Synovial Fluid of Rheumatoid Arthritis Patient

Abstract

This study included the isolation of the enzyme Matrix Metalloprotrinase-1 (MMP-1; Collagenase-1) from serum of healthy individual and synovial fluid (SF) of a patient with rheumatoid arthritis using different techniques. After precipitation of proteins using saturated ammonium sulfate, two porteinous components had been isolated by gel filtration chromatography. It was found that only the first peak has high activity of latent-MMP-1. The apparent molecular weights of MMP-1 in serum and SF using gel filtration chromatography was found to be (47752 ± 816 and 48194 ± 707) dalton respectively. High performance liquid chromatography (HPLC) was used to show the extent of purity. The main maximum of the enzyme from serum and SF appeared at retention times (1.652 and 1.65) minutes respectively compared with the retention time of standard enzyme at (1.571) minutes. The approximate molecular weight of latent-MMP-1 by HPLC technique, in serum and SF were found (48067 and 48033) dalton respectively. The study included, also, the effect of some material compounds on the activity of latent-MMP-1. The results revealed that the addition of ethylene diamine tetra acetic acid (EDTA), 2,4-dinitrophenol, sodium azide, potassium oxalate, mercaptoethanol (MEH), glutathione (GSH), cystein and healthy human serum decreased the activity of latent-MMP-1, while the addition of NaCl, MgSO 4 , CoCl 2 , CaCl 2 , CuSO 4 , ZnSO 4 , HgCl 2 and pepsin increases the activity of enzyme.

Adenosine Deaminase Activity a Biomarker for Rheumatoid Arthritis: Review

“…The synovial tissue in RA is described by prominent hyperplasia, called pannus, which contains pro-inflammatory cytokines [36,63,64] and extents over and under the articular cartilage [62]. In the affected joints, many substances such as tumor necrosis factor (TNF), interleukin-1 (IL-1), fibroblasts, metalloproteinases (MMPs), aggrecanases, and cathepsins are produced [22,54,65,66].…”