MMP 8 Human, His
Description
Production and Purification
MMP-8 Human, His is typically produced in E. coli or mammalian systems . Key steps include:
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Expression: Recombinant synthesis in bacterial systems (e.g., E. coli) yields non-glycosylated protein .
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Purification: Nickel/cobalt affinity chromatography leverages the His-tag .
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Activation: Proenzyme requires activation via 1 mM APMA (4-aminophenylmercuric acetate) at 37°C for 1 hour .
Example Protocol (Activation) :
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Dissolve rhMMP-8 at 100 µg/mL in assay buffer (50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, pH 7.5).
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Add 1 mM APMA (100 mM stock in DMSO).
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Incubate at 37°C for 1 hour.
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Dilute to 1.0 ng/µL for enzymatic assays.
Enzymatic Activity
MMP-8 cleaves collagen substrates with high specificity. Activity is quantified using FRET-based assays with substrates like MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂ .
| Parameter | Value | Source |
|---|---|---|
| Specific Activity | >250 pmol/min/µg (activated) | |
| Substrate | MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂ (2 mM) | |
| Optimal pH | 7.5 (assay buffer) |
Functional Domains
Inflammatory Processes
MMP-8 modulates proinflammatory cytokines and chemokines:
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TNF-α Processing: Cleaves pro-TNF-α to release active TNF-α, enhancing inflammation .
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Neutrophil Recruitment: Activates CXCL8/IL-8 and CXCL5, amplifying neutrophil chemotaxis .
Cardiovascular Diseases
MMP-8 is implicated in atherosclerosis:
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Collagen Degradation: Degrades type I collagen in atheroma, contributing to plaque instability .
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Cellular Expression: Found in endothelial cells, smooth muscle cells, and macrophages within atherosclerotic lesions .
Cancer Biology
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Metastasis Suppression: Inhibits tumor invasion in breast and oral squamous cell carcinomas (OTSCC) .
Key Research Findings
Challenges and Considerations
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Glycosylation: Native MMP-8’s glycosylation enhances stability; recombinant forms (e.g., E. coli) lack this feature, potentially altering activity .
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Activation Dependency: Proenzyme requires APMA or oxidative agents (e.g., hypochlorous acid) for activation .
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Storage: Avoid repeated freeze-thaw cycles; store at -20°C for long-term stability .
Product Specs
Product Science Overview
Matrix Metalloproteinase-8 (MMP-8), also known as neutrophil collagenase, is a member of the matrix metalloproteinase (MMP) family. These enzymes are involved in the breakdown of extracellular matrix components, playing crucial roles in various physiological and pathological processes. MMP-8 is particularly known for its ability to degrade type I, II, and III collagens .
MMP-8 plays a significant role in normal physiological processes such as embryonic development, reproduction, and tissue remodeling. It is also involved in disease processes, including arthritis and metastasis . In cancer research, the loss of MMP-8 has been associated with increased tumor growth and metastatic burden, as well as enhanced tumor vascularity and altered immune cell infiltration .
Recombinant human MMP-8 with a His tag is a form of the protein that has been genetically engineered for research purposes. The His tag is a sequence of histidine residues added to the protein to facilitate purification and detection. This recombinant protein is typically expressed in systems such as HEK 293 cells or E. coli .
