MMP 3 Human, HEK

Matrix Metalloproteinase-3 Human Recombinant, HEK
Cat. No.
BT8202
Source
HEK293 cells.
Synonyms
Stromelysin-1, EC 3.4.24.17, Matrix metalloproteinase-3, MMP-3, Transin-1, SL-1, STMY, STR1, STMY1, MGC126102, MGC126103, MGC126104.
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 95% as determined by SDS-PAGE.
Usage
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

MMP-3 Human Recombinant produced in HEK293 cells is a proform of the Human MMP3 [Tyr18-Cys477 (Lys45Glu)] and fused with a ployhistide tag at the C-terminus, having an Mw of 52kDa.
MMP-3 is purified by proprietary chromatographic techniques.

Product Specs

Introduction
MMP-3, also known as Stromelysin-1 or Transin-1, is an enzyme that breaks down natural collagen at normal body pH and temperature. It plays a role in the breakdown of the intervertebral disc, specifically the nucleus pulposus and annulus fibrosus, in cases of Herniated Lumbar Intervertebral Disk. MMP-3 breaks down various components of the extracellular matrix, including proteoglycan, laminin, fibronectin, gelatin, and collagen types III, IV, and IX. It also activates pro-MMP-9 and pro-MMP-8, and enhances the activation of plasmin-activated MMP-1. MMP-3 is initially produced in an inactive form (proenzyme) and is activated by various proteinases such as plasmin, trypsin, chymotrypsin, cathepsin G, and human neutrophil elastase. Furthermore, MMP-3 has been shown to activate the precursor of IL1-beta.
Description
Recombinant Human MMP-3, produced in HEK293 cells, is a proform of human MMP3 [Tyr18-Cys477 (Lys45Glu)] with a polyhistidine tag at the C-terminus, resulting in a molecular weight of 52kDa. This protein is purified using proprietary chromatographic techniques.
Physical Appearance
A clear, colorless solution that has been sterilized by filtration.
Formulation
MMP-3 is provided as a 0.2µm filtered solution in a buffer containing 20mM Tris-HCl, 150mM NaCl, and 0.05% Brij35, at pH 7.5.
Stability
For short-term storage (up to 2-4 weeks), keep at 4°C. For long-term storage, freeze at -20°C. Repeated freezing and thawing should be avoided.
Purity
Purity is determined to be greater than 95% using SDS-PAGE analysis.
Biological Activity
The enzyme's activity is assessed by its ability to break down the fluorescent peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2. A specific activity greater than 150 pmoles/min/µg is observed. Recombinant Human MMP-3 protein pro form requires activation with Chymotrypsin. Here's the activation protocol: 1) Dilute MMP3 to 20µg/ml in Assay Buffer (50mM Tris, 10mM CaCl2, 150mM NaCl, 0.05% (w/v) Brij 35, pH 7.5). 2) Activate MMP3 by adding Chymotrypsin (Sigma, Catalog#C­3142, 1mg/ml stock in 1mM HCl) to a final concentration of 5µg/ml. 3) Incubate at 37°C for 30 minutes. 4) Stop the activation by adding 2mM PMSF (pre-warmed to 37°C).
Synonyms
Stromelysin-1, EC 3.4.24.17, Matrix metalloproteinase-3, MMP-3, Transin-1, SL-1, STMY, STR1, STMY1, MGC126102, MGC126103, MGC126104.
Source
HEK293 cells.

Product Science Overview

Structure and Function

MMP-3 is synthesized as an inactive proenzyme (zymogen) and requires activation to become functionally active. The human recombinant form of MMP-3 is produced in HEK293 cells (Human Embryonic Kidney 293 cells), which are commonly used in biotechnology for the production of recombinant proteins . The recombinant MMP-3 is typically fused with a polyhistidine tag at the C-terminus to facilitate purification .

The active form of MMP-3 can degrade a wide range of ECM components, including proteoglycans, laminin, fibronectin, and collagen . Additionally, MMP-3 can activate other pro-MMPs, such as MMP-1, MMP-7, and MMP-9, thereby amplifying the ECM degradation process .

Biological Significance

MMP-3 plays a significant role in various biological processes:

  1. Tissue Remodeling: MMP-3 is involved in the normal turnover and remodeling of the ECM, which is essential for tissue repair and regeneration.
  2. Wound Healing: MMP-3 contributes to the wound healing process by breaking down damaged ECM components and facilitating the migration of cells to the wound site.
  3. Inflammation: MMP-3 is implicated in inflammatory processes, where it can modulate the activity of cytokines and chemokines.
  4. Cancer Metastasis: MMP-3 is associated with cancer progression and metastasis by degrading ECM barriers and promoting tumor cell invasion.
Clinical Relevance

Due to its involvement in various pathological conditions, MMP-3 is a potential therapeutic target for diseases such as arthritis, cardiovascular diseases, and cancer. Inhibitors of MMP-3 are being explored for their potential to modulate ECM degradation and reduce tissue damage in these diseases.

Production and Purification

The human recombinant MMP-3 produced in HEK293 cells is purified using proprietary chromatographic techniques to achieve high purity levels (greater than 95% as determined by SDS-PAGE) . The protein is supplied as a sterile filtered colorless solution in a specific formulation to maintain its stability and activity .

Storage and Stability

The recombinant MMP-3 should be stored at 4°C if it will be used within 2-4 weeks. For longer storage periods, it should be kept frozen at -20°C to maintain its stability. It is important to avoid multiple freeze-thaw cycles to prevent degradation of the protein .

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