TIMP2 Human, His
Description
Introduction to TIMP2 Human, His
TIMP2 Human, His refers to recombinant human tissue inhibitor of metalloproteinase-2 (TIMP-2) engineered with a C-terminal polyhistidine (His) tag for enhanced purification and detection. Produced in human embryonic kidney (HEK-293) cells, this protein retains its native structure and function while enabling efficient bioprocessing . TIMP-2 is a 21–22 kDa glycoprotein that regulates extracellular matrix (ECM) remodeling by inhibiting matrix metalloproteinases (MMPs), particularly MMP-2, and modulating MMP activation through interactions with membrane-type MMPs (MT-MMPs) . The His-tag facilitates affinity chromatography purification, yielding high-purity preparations (>95%) critical for biochemical and therapeutic applications .
Production and Purification
TIMP2 Human, His is synthesized via transient or stable expression in HEK-293 cells, leveraging codon optimization for enhanced yield (>35 mg/L in optimized cultures) . Key steps include:
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Expression: TIMP-2 cDNA (Cys27-Pro220) is transfected into HEK-293 cells, with secretion into the culture medium .
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Purification:
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Quality Control:
Biological Functions
TIMP2 Human, His exhibits dual roles in ECM and cellular regulation:
MMP Inhibition and Activation
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MMP-2 Inhibition: Directly binds active MMP-2, blocking ECM degradation (IC₅₀ ≈ 2.2 nM) .
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MMP-2 Activation: Forms a complex with pro-MMP-2, enabling MT1-MMP-mediated activation at the cell surface .
Non-MMP Functions
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Endothelial Cell Suppression: Inhibits angiogenesis independently of MMP inhibition .
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Neuronal Plasticity: Regulates hippocampal ECM remodeling, dendritic spine density, and adult neurogenesis .
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Growth Promotion: Stimulates cell proliferation via receptor-mediated signaling (e.g., binding to high-affinity sites on Raji cells) .
Cardiovascular and Neurological Diseases
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Myocardial Infarction: TIMP2 deficiency exacerbates LV dilation and dysfunction post-MI due to unchecked MT1-MMP activity .
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Cognitive Decline: TIMP2-IgG4 fusion proteins restore hippocampal function and memory in aged mice, independent of MMP inhibition .
Cancer and ECM Pathologies
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Tumor Growth: Inhibits angiogenesis and metastasis by suppressing MMP-2 activation and endothelial proliferation .
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Atherosclerosis: Reduces plaque destabilization by limiting macrophage migration and ECM breakdown .
Engineered Variants
Therapeutic Potential and Challenges
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Opportunities:
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Challenges:
Product Specs
Product Science Overview
Tissue Inhibitor of Metalloprotease 2 (TIMP-2) is a secreted, multifunctional protein with a molecular weight of approximately 21 kDa . It is a member of the TIMP family, which includes four endogenous proteins that primarily function to inhibit the activities of matrix metalloproteinases (MMPs) . TIMP-2 is widely expressed in mammalian tissues and plays a crucial role in maintaining the balance between proteases and their inhibitors, which is essential for the regulation of extracellular matrix (ECM) turnover .
The human recombinant TIMP-2 with a His tag is typically produced using prokaryotic expression systems. The gene encoding TIMP-2 is cloned into a suitable expression vector, such as pET28a, and transformed into Escherichia coli (E. coli) BL21 (DE3) cells . The expression of TIMP-2 is induced by isopropyl-β-D-thiogalactoside (IPTG), and the protein is subsequently purified using nickel affinity chromatography . The His tag facilitates the purification process by allowing the recombinant protein to bind to the nickel ions on the affinity column .
TIMP-2 is known for its ability to inhibit the activity of MMPs, particularly MMP-2 . MMPs are a family of enzymes responsible for the degradation of the ECM, which is a critical process in various physiological and pathological conditions, including tissue remodeling, wound healing, and tumor invasion . By inhibiting MMPs, TIMP-2 helps to regulate ECM turnover and maintain tissue integrity .
In addition to its role as an MMP inhibitor, TIMP-2 is also involved in the activation of pro-MMP-2 through its interaction with MMP-14 . This dual function of TIMP-2 highlights its importance in the fine-tuning of ECM remodeling processes .
TIMP-2 is ubiquitously expressed in various tissues, including the lungs, liver, kidneys, and heart . Its expression levels can vary depending on the tissue type and the physiological or pathological state of the organism . For example, elevated levels of TIMP-2 have been observed in fibrotic tissues, where it contributes to ECM accumulation and fibrosis .
The expression of TIMP-2 is tightly regulated at both the transcriptional and post-transcriptional levels. Various cytokines, growth factors, and hormones can modulate TIMP-2 expression . Additionally, microRNAs, such as miR-410, have been shown to negatively regulate TIMP-2 expression, which is associated with the progression of non-small-cell lung cancer .
TIMP-2 has significant clinical implications, particularly in the context of cancer and fibrotic diseases. Its ability to inhibit MMPs makes it a potential therapeutic target for preventing tumor invasion and metastasis . Moreover, the regulation of TIMP-2 expression and activity could be a promising strategy for managing fibrotic conditions .
