MIP 3a Human, His
Description
Solution NMR and Crystallographic Structures
MIP-3α adopts the canonical chemokine fold:
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Three antiparallel β strands (β1: residues 20–26, β2: 36–41, β3: 46–49)
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C-terminal α helix (residues 54–63) at 69° relative to β strands (solution NMR) .
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C-terminal peptide (residues 51–70) forms an amphipathic α helix critical for antimicrobial activity .
Key Differences Between Monomeric and Dimeric Forms
Primary Receptor and Immune Recruitment
MIP-3α binds CCR6 (its exclusive receptor) to recruit:
Antimicrobial Properties
The C-terminal α-helical region exhibits broad-spectrum activity:
| Target Organism | Activity | Source |
|---|---|---|
| Escherichia coli | Strong bactericidal | |
| Staphylococcus aureus | Moderate bactericidal | |
| HIV-1 | Host restriction via CCR6 activation |
Role in Human Pathologies
Experimental Models
Product Specs
Q&A
Experimental Design: How to Optimize MIP-3α Expression in E. coli?
When expressing MIP-3α in Escherichia coli, optimizing conditions is crucial for high yield and purity. Consider the following steps:
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Temperature and Induction: Optimize temperature (e.g., 25°C) and induction conditions (e.g., IPTG concentration) to enhance protein expression.
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Media Composition: Use rich media (e.g., LB or TB) supplemented with necessary nutrients to support bacterial growth.
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Purification Techniques: Employ efficient chromatographic methods (e.g., Ni-NTA for His-tagged proteins) to purify the recombinant protein .
Structural Analysis: What Are the Key Differences Between Monomeric and Dimeric MIP-3α?
MIP-3α can exist as both monomers and dimers, with distinct structural features:
| Feature | Monomeric MIP-3α | Dimeric MIP-3α |
|---|---|---|
| Binding Groove Width | Larger (~13.8 Å) | Smaller (~12.1 Å) |
| C-terminal α Helix Stability | More flexible | More stable |
| pH Dependence | Predominantly monomeric at acidic pH | Dimerization favored at neutral pH |
These differences can affect receptor binding and biological activity .
Data Contradiction Analysis: How to Resolve Discrepancies in MIP-3α Structural Studies?
Discrepancies between NMR and crystal structures of MIP-3α can arise from differences in experimental conditions:
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NMR vs. Crystallography: NMR structures often reflect solution conditions, while crystal structures may impose constraints not present in solution.
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pH and Ionic Strength: These factors can influence protein conformation and oligomerization state.
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Species-Specific Variations: Differences between human and murine MIP-3α structures highlight the importance of species-specific sequences .
Biological Function: What Role Does MIP-3α Play in Immune Response and Disease?
MIP-3α (CCL20) is crucial for recruiting immune cells to sites of inflammation:
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Immune Cell Recruitment: Attracts memory T cells, natural killer cells, and immature dendritic cells via CCR6.
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Disease Association: Linked to conditions like rheumatoid arthritis, cancer, and periodontal diseases.
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Antimicrobial Activity: Exhibits direct antimicrobial properties, contributing to its role in infection control .
Methodological Approach: How to Measure MIP-3α Concentrations in Biological Samples?
To quantify MIP-3α in biological samples, use ELISA kits:
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ELISA Kits: Utilize kits like the Human MIP-3 alpha ELISA Kit for precise measurement.
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Sample Preparation: Ensure proper dilution and handling of samples to avoid interference.
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Data Analysis: Plot standard curves to interpolate concentrations from sample readings .
Advanced Experimental Design: How to Investigate the Role of MIP-3α in Specific Disease Models?
To study MIP-3α’s role in disease models:
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In Vitro Models: Use cell cultures (e.g., PBMCs or THP-1 cells) stimulated with appropriate factors to mimic disease conditions.
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In Vivo Models: Employ animal models of diseases (e.g., arthritis or cancer) to assess MIP-3α’s effects on disease progression.
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Data Analysis: Compare outcomes between control and MIP-3α-treated groups to elucidate its role .
Product Science Overview
CCL20 is expressed in various tissues, including the liver, lungs, lymph nodes, and peripheral blood lymphocytes . Its expression is strongly upregulated by inflammatory signals and downregulated by the anti-inflammatory cytokine IL-10 . This regulation ensures that CCL20 is produced in response to inflammation, aiding in the recruitment of immune cells to the site of infection or injury.
CCL20 exerts its effects by binding to the G protein-coupled receptor CCR6 . This interaction is crucial for the chemoattraction of lymphocytes and dendritic cells towards epithelial cells, particularly in mucosal tissues . The protein is strongly chemotactic for lymphocytes and weakly attracts neutrophils . It is also involved in the formation and function of mucosal lymphoid tissues, which are essential for the body’s immune defense .
Recombinant human CCL20 is a non-glycosylated protein consisting of 70 amino acids and has a molecular weight of approximately 8.0 kDa . The recombinant form is often tagged with a His (histidine) tag to facilitate purification and detection in research applications. This form is widely used in scientific studies to understand the protein’s function and its role in various biological processes.
Recombinant CCL20 is primarily used for research purposes, including studies on immune response, inflammation, and cell signaling. It is not intended for human, animal, or diagnostic applications . Researchers utilize this protein to investigate its role in attracting immune cells and its potential therapeutic applications in treating inflammatory diseases.
