GO:0035715 chemokine (C-C motif) ligand 2 binding: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0035715 (chemokine (C-C motif) ligand 2 binding) is a molecular function term describing the selective interaction of a protein with CCL2, a CC-chemokine also known as MCP-1.
• CCL2 binding is central to immune cell recruitment, tumor microenvironment remodeling, and inflammatory signaling in cancer, metabolic disease, and neuroinflammation.
• The CCL2-CCR2 axis is the best-characterized downstream signaling pathway triggered by CCL2 binding, but GO:0035715 specifically captures the binding event itself, not receptor activation.
• Key proteins annotated with CCL2 binding include CCR2, ACKR2/D6, and glycosaminoglycan-presenting proteins that immobilize CCL2 on endothelial surfaces.
• Dysregulated CCL2 binding contributes to glioblastoma immunosuppression, esophageal squamous cell carcinoma progression, diabetic anxiety, and MASLD.
• CRISPR knockout, knock-in, and overexpression models are essential to dissect which domains of CCL2-binding proteins mediate functional interactions in vivo.
Description
GO:0035715, chemokine (C-C motif) ligand 2 binding, is a molecular function term in the Gene Ontology that describes the selective interaction between a protein and CCL2 (also known as MCP-1), a CC-chemokine that orchestrates monocyte, macrophage, and T-cell trafficking. CCL2 is one of the most extensively studied chemokines because of its dual roles in host defense and in pathological inflammation, including cancer, obesity, and neurodegeneration. The binding event captured by GO:0035715 is the first committed step in CCL2 biology, preceding receptor activation, gradient formation, and downstream signaling. Researchers study GO:0035715 because it defines which proteins physically engage CCL2 and therefore which cells sense, sequester, or present this chemokine. For example, CCR2 is the canonical signaling receptor for CCL2, while ACKR2 acts as a scavenging receptor that internalizes CCL2 to shape chemokine gradients. In the tumor microenvironment, CCL2 binding to CCR2 on monocytes and tumor-associated macrophages promotes immunosuppression and tumor progression in glioblastoma and esophageal squamous cell carcinoma. In metabolic tissues, CCL2 binding influences adipogenesis, angiogenesis, and lipid accumulation, linking this molecular function to obesity, diabetes, and steatotic liver disease. Because CCL2 binding is a molecular function rather than a single pathway, it intersects with diverse biological processes: embryo implantation in the bovine endometrium, IgE-mediated mast cell activation, and neuronal stress responses in diabetes. Understanding the structural and cellular context of CCL2 binding is therefore essential for designing therapeutics that selectively block pathogenic CCL2 interactions without compromising host immunity.
chemokine (C-C motif) ligand 2 binding At A Glance
| GO ID | GO:0035715 |
|---|---|
| GO term | chemokine (C-C motif) ligand 2 binding |
| Ontology | molecular_function |
| Synonym | CCL2 binding |
| Definition | Binding to chemokine (C-C motif) ligand 2. |
| Major function | Mediates selective recognition of CCL2 by receptors, decoy receptors, and matrix proteins, initiating or modulating CCL2-dependent signaling. |
| Related ligand | CCL2 (MCP-1), a CC-chemokine that recruits monocytes, macrophages, and T cells. |
| Canonical receptor | CCR2, a G-protein-coupled receptor that signals upon CCL2 binding. |
| Scavenging receptor | ACKR2 (D6), a atypical chemokine receptor that binds and internalizes CCL2. |
| Disease relevance | Cancer, metabolic dysfunction-associated steatotic liver disease, diabetic anxiety, and implantation failure. |
What Is GO:0035715?
GO:0035715, chemokine (C-C motif) ligand 2 binding, is defined as the selective and non-covalent interaction of a protein with chemokine (C-C motif) ligand 2 (CCL2). This molecular function encompasses the physical association between CCL2 and any protein that recognizes it, including signaling receptors, decoy receptors, and extracellular matrix components that immobilize the chemokine. The term does not describe downstream signaling events; it captures the binding event itself, which is a prerequisite for CCL2-mediated cellular responses such as chemotaxis, integrin activation, and inflammatory gene expression.
Why Is chemokine (C-C motif) ligand 2 binding Important in Cell Biology?
GO:0035715 is important because CCL2 binding is a nodal point in inflammatory and homeostatic signaling that determines whether tissues recruit immune cells, sequester chemokines, or mount pathological inflammatory responses. Dysregulated CCL2 binding is implicated in tumor immune evasion, metabolic inflammation, and neuropsychiatric complications of diabetes, making it a high-value target for therapeutic intervention and biomarker discovery. Understanding the molecular determinants of CCL2 binding also informs the design of biologics and small molecules that selectively disrupt pathogenic interactions while preserving beneficial chemokine functions.
• CCL2 binding to CCR2 on monocytes and macrophages is a driver of tumor-associated macrophage recruitment and immunosuppression in glioblastoma.
• In esophageal squamous cell carcinoma, the LINC00330/CCL2 axis reprograms tumor-associated macrophages and promotes tumor progression.
• CCL2 binding regulates prostaglandin synthesis and embryo attachment in the bovine endometrium during implantation.
• Neuronal CCL2 binding contributes to anxiety disorders in the context of diabetes and hyperglycaemia.
• CCL2 binding to CCR2 promotes lipid accumulation in metabolic dysfunction-associated steatotic liver disease.
• CCL2 binding enhances adipogenesis and angiogenesis in adipose tissue engineering co-culture systems.
• CCL2 binding is a key step in IgE-mediated FcεRI activation and mast cell biology.
• CCL2 binding promotes prostate cancer growth through multiple mechanisms, including angiogenesis and immune evasion.
• The CCL2-CCR2 axis is a validated therapeutic target, and blocking CCL2 binding is an active area of drug development.
• GO:0035715 provides a precise annotation for functional studies of CCL2-interacting proteins in health and disease.
Molecular Mechanism of chemokine (C-C motif) ligand 2 binding
CCL2 Recognition by CCR2
In simple terms: CCL2 docks onto CCR2 like a key in a lock, triggering signals that call immune cells into tissues.
The best-characterized CCL2 binding event is its interaction with CCR2, a seven-transmembrane G-protein-coupled receptor expressed on monocytes, macrophages, and some tumor cells. Structural and functional studies indicate that CCL2 binds CCR2 through a two-step mechanism involving the chemokine N-terminus and the receptor extracellular loops, leading to G-protein activation and downstream calcium flux, PI3K/AKT signaling, and integrin activation. This binding event is essential for monocyte recruitment in inflammation and for tumor-associated macrophage polarization in cancer.
Scavenging and Gradient Regulation by ACKR2
In simple terms: ACKR2 acts like a sponge that soaks up CCL2, shaping the chemical trail that guides immune cells.
ACKR2 (also known as D6) is an atypical chemokine receptor that binds CCL2 and other inflammatory CC-chemokines with high affinity but does not signal through G-proteins. Instead, ACKR2 internalizes CCL2 and targets it for degradation, thereby shaping chemokine gradients and limiting excessive inflammation. This scavenging function is critical in resolving inflammation and preventing tissue damage, and its dysregulation has been linked to cancer progression and chronic inflammatory diseases.
Extracellular Matrix and Glycosaminoglycan Presentation
In simple terms: CCL2 sticks to sugar chains on cell surfaces and matrix, creating a local concentration that immune cells can follow.
CCL2 binding to glycosaminoglycans (GAGs) on endothelial cells and extracellular matrix components is essential for immobilizing the chemokine and presenting it to rolling leukocytes. This interaction creates haptotactic gradients that guide cell migration and is required for CCL2-mediated angiogenesis and adipogenesis in tissue engineering models. The GAG-binding site on CCL2 is distinct from the receptor-binding site, allowing simultaneous presentation and receptor engagement.
Regulation of CCL2 Binding by Post-Translational Modifications
In simple terms: Chemical tags on CCL2 or its binding partners can switch interactions on or off.
CCL2 binding can be modulated by post-translational modifications, including glycosylation and proteolytic processing of the chemokine N-terminus. In the bovine endometrium, CCL2 regulates prostaglandin synthesis and embryo attachment, and its binding activity is influenced by the local hormonal milieu. In mast cells, IgE-mediated FcεRI activation induces CCL2 expression and binding to receptors that amplify allergic inflammation. These examples illustrate that CCL2 binding is not constitutive but is dynamically regulated by tissue context and inflammatory cues.
Pathological Consequences of Dysregulated CCL2 Binding
In simple terms: When CCL2 binding goes wrong, it can fuel cancer, metabolic disease, and nerve damage.
Dysregulated CCL2 binding contributes to multiple pathologies. In glioblastoma, Lnc-H19-derived protein shapes an immunosuppressive microenvironment in part through CCL2-dependent mechanisms. In esophageal squamous cell carcinoma, the LINC00330/CCL2 axis reprograms tumor-associated macrophages to promote progression. In metabolic dysfunction-associated steatotic liver disease, CCL2 binding to CCR2 promotes lipid accumulation and inflammation. In diabetes, neuronal CCL2 binding contributes to anxiety disorders. These findings underscore the importance of precise regulation of GO:0035715 in human health.
Key Genes Involved in GO:0035715 chemokine (C-C motif) ligand 2 binding
The following genes and proteins are directly or indirectly involved in chemokine (C-C motif) ligand 2 binding (GO:0035715) and its downstream biology.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CCL2 | Ligand that binds CCR2 and other partners; also known as MCP-1 | Central to monocyte recruitment, tumor microenvironment remodeling, and metabolic inflammation |
| CCR2 | Canonical signaling receptor for CCL2 | Mediates CCL2-dependent chemotaxis and is a therapeutic target in cancer and fibrosis |
| ACKR2 | Atypical scavenging receptor that binds and internalizes CCL2 | Regulates chemokine gradients and inflammation resolution |
| LINC00330 | Long non-coding RNA that modulates CCL2 expression in ESCC | Reprograms tumor-associated macrophages via the CCL2 axis |
| Lnc-H19 | Long non-coding RNA that encodes a protein shaping glioblastoma microenvironment | Links CCL2 binding to immunosuppression in glioblastoma |
| FcεRI | High-affinity IgE receptor on mast cells | IgE-mediated activation induces CCL2 binding and allergic inflammation |
| GAGs | Glycosaminoglycans on cell surfaces and matrix | Present CCL2 to leukocytes and modulate angiogenesis |
| CCR2B | Splice variant of CCR2 | Isoform-specific CCL2 binding in monocytes |
| MCP-1 | Alternative name for CCL2 | Used interchangeably in literature on CCL2 binding |
| TAMs | Tumor-associated macrophages | CCL2 binding promotes their recruitment and immunosuppressive phenotype |
| HUVECs | Human umbilical vein endothelial cells | Model system for CCL2-mediated angiogenesis |
| ADSCs | Adipose-derived stem cells | CCL2 binding enhances adipogenesis in co-culture |
| Prostaglandin synthase | Enzyme induced by CCL2 in endometrium | CCL2 binding regulates embryo implantation |
| Neuronal CCL2 | CCL2 expressed by neurons under hyperglycaemia | Contributes to anxiety in diabetes |
| CCR2 antagonists | Small molecules blocking CCL2-CCR2 binding | Therapeutic candidates for MASLD and cancer |
| 2,5-DHBA | Natural compound targeting CCL2-CCR2 axis | Ameliorates MASLD by reducing lipid accumulation |
| IL-6 | Cytokine induced by CCL2 binding | Amplifies inflammatory signaling in tumors |
| TGF-β | Cytokine modulated by CCL2 binding | Promotes immunosuppression in glioblastoma |
How Is chemokine (C-C motif) ligand 2 binding Regulated?
CCL2 binding is regulated at multiple levels. Transcriptional regulation of CCL2 itself is controlled by NF-κB and STAT pathways in response to inflammatory stimuli. Post-translational modifications, including glycosylation and proteolytic cleavage, can alter CCL2's binding affinity for CCR2 and GAGs. Receptor availability is regulated by internalization and recycling of CCR2 and ACKR2. In metabolic tissues, hyperglycaemia and lipid overload increase CCL2 expression and binding, contributing to diabetic complications and steatotic liver disease. Therapeutic interventions that target the CCL2-CCR2 axis, such as 2,5-dihydroxybenzoic acid, can modulate CCL2 binding and reduce lipid accumulation in MASLD models.
chemokine (C-C motif) ligand 2 binding and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CCL2 | Glioblastoma immunosuppression | U251 or U87 glioblastoma xenografts with CCL2 knockout |
| CCR2 | Esophageal squamous cell carcinoma progression | KYSE cell lines co-cultured with macrophages |
| CCL2 | Metabolic dysfunction-associated steatotic liver disease | High-fat diet mouse model treated with CCL2-CCR2 inhibitors |
| CCL2 | Diabetic anxiety disorders | Streptozotocin-induced diabetic mice with neuronal CCL2 knockdown |
| CCL2 | Bovine embryo implantation failure | Bovine endometrial epithelial cell cultures |
CCL2 Binding in Cancer Progression and Immunosuppression
CCL2 binding to CCR2 on monocytes and tumor-associated macrophages promotes an immunosuppressive tumor microenvironment in glioblastoma and esophageal squamous cell carcinoma. In glioblastoma, Lnc-H19-derived protein shapes this microenvironment, and CCL2 binding is a key mediator of macrophage recruitment. In ESCC, the LINC00330/CCL2 axis reprograms TAMs to support tumor progression. In prostate cancer, CCL2 binding promotes tumor growth through multiple mechanisms, including angiogenesis and survival signaling. These findings establish CCL2 binding as a therapeutic target in oncology.
CCL2 Binding in Metabolic and Liver Disease
CCL2 binding to CCR2 contributes to lipid accumulation and inflammation in metabolic dysfunction-associated steatotic liver disease (MASLD). Targeting the CCL2-CCR2 axis with 2,5-dihydroxybenzoic acid ameliorates MASLD in preclinical models. In adipose tissue, CCL2 binding enhances adipogenesis and angiogenesis, linking this molecular function to obesity and metabolic syndrome. These studies highlight the potential of modulating CCL2 binding for metabolic therapeutics.
CCL2 Binding in Neuroinflammation and Psychiatric Disorders
Neuronal CCL2 responds to hyperglycaemia and contributes to anxiety disorders in the context of diabetes. This suggests that CCL2 binding in the central nervous system is not merely a marker of inflammation but an active driver of behavioral changes. Understanding how CCL2 binding is regulated in neurons may open new avenues for treating diabetes-associated mood disorders.
CCL2 Binding in Reproductive and Allergic Biology
In the bovine endometrium, CCL2 regulates prostaglandin synthesis and embryo attachment during implantation, indicating a role for CCL2 binding in reproductive success. In mast cells, IgE-mediated FcεRI activation induces CCL2 binding, linking this molecular function to allergic inflammation. These diverse contexts demonstrate the broad biological significance of GO:0035715.
From chemokine (C-C motif) ligand 2 binding-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does CCL2 binding to CCR2 drive monocyte recruitment in tumors? | CCR2 knockout mice with syngeneic tumor implants |
| Which domains of CCL2 mediate GAG binding versus receptor binding? | Point-mutant CCL2 knock-in mice |
| Can blocking CCL2 binding reverse MASLD? | Liver-specific CCL2 overexpression or knockout in diet-induced obesity models |
| Does neuronal CCL2 binding cause anxiety in diabetes? | Neuron-specific CCL2 knockout mice under hyperglycaemia |
| How does ACKR2 scavenging affect CCL2 gradients? | ACKR2 knockout zebrafish or mouse models |
| Does CCL2 binding regulate embryo implantation? | Bovine endometrial explants with CCL2 knockdown |
How to Study the chemokine (C-C motif) ligand 2 binding Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Surface plasmon resonance | Binding affinity and kinetics | CCL2-CCR2 interaction studies |
| Isothermal titration calorimetry | Thermodynamics of binding | CCL2-GAG interactions |
| Calcium flux assay | G-protein activation | CCR2 signaling in monocytes |
| Chemotaxis assay | Cell migration | Monocyte recruitment in vitro |
| RNA-seq | Transcriptional changes | Tumor microenvironment profiling |
| Proteomics | Protein expression and interactions | CCL2 binding partners identification |
| CRISPR knockout | Gene function | CCR2 and ACKR2 loss-of-function studies |
| In vivo tumor models | Tumor growth and immune infiltration | Glioblastoma and ESCC xenografts |
Binding Assays for CCL2-Protein Interactions
Surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) are used to measure the affinity and kinetics of CCL2 binding to CCR2, ACKR2, and GAGs. These biophysical methods provide quantitative parameters such as KD and stoichiometry, which are essential for understanding the molecular basis of GO:0035715.
Cell-Based Signaling Assays
Calcium flux assays, cAMP inhibition assays, and chemotaxis assays are used to measure functional consequences of CCL2 binding to CCR2 in monocytes and macrophages. These assays can be combined with CRISPR knockout of CCR2 or ACKR2 to dissect receptor-specific contributions.
Transcriptomic and Proteomic Profiling
RNA-seq and proteomics are used to identify genes and proteins whose expression is altered by CCL2 binding in tumor cells, endothelial cells, and immune cells. These approaches reveal downstream pathways and potential therapeutic targets.
In Vivo Models of CCL2 Binding
Mouse models of cancer, obesity, diabetes, and liver disease are used to study the role of CCL2 binding in vivo. Genetic knockout of CCL2 or CCR2, or pharmacological blockade of CCL2 binding, can be used to assess therapeutic potential.
How CRISPR Can Be Used to Study GO:0035715 chemokine (C-C motif) ligand 2 binding
Knockout
CRISPR knockout of CCR2 or ACKR2 is used to determine which receptor mediates specific CCL2 binding functions in immune cells and tumors. Knockout of CCL2 itself in cancer cell lines can reduce macrophage recruitment and tumor growth in xenograft models.
Point Mutation
Point mutations in the CCL2-binding interface of CCR2 or in the GAG-binding site of CCL2 can be introduced using CRISPR base editing or homology-directed repair to dissect binding specificity. These models are valuable for separating signaling from scavenging functions.
Knock-in
Knock-in of tagged CCL2 or CCR2 (e.g., GFP or HA) allows visualization and immunoprecipitation of CCL2-binding complexes in live cells and tissues. This approach is useful for mapping CCL2 binding sites in vivo.
Overexpression
Overexpression of CCL2 or CCR2 in cell lines and mouse models is used to study gain-of-function effects on inflammation, angiogenesis, and tumor progression. Overexpression models can also be used to test CCL2-binding inhibitors.
How EDITGENE Supports chemokine (C-C motif) ligand 2 binding Research
Researchers studying chemokine (C-C motif) ligand 2 binding-related genes often need to determine whether a candidate gene is causally involved in CCL2 recognition, signaling, or downstream pathology. EDITGENE provides end-to-end CRISPR solutions to generate precisely engineered cell and animal models, enabling rigorous functional validation of GO:0035715-associated genes.
Contact EDITGENE today to design your custom CRISPR model for chemokine (C-C motif) ligand 2 binding research.
Frequently Asked Questions About chemokine (C-C motif) ligand 2 binding
What is GO:0035715?
GO:0035715 is the Gene Ontology molecular function term for chemokine (C-C motif) ligand 2 binding, describing the selective interaction of a protein with CCL2.
What genes are involved in chemokine (C-C motif) ligand 2 binding?
Key genes include CCL2, CCR2, ACKR2, and LINC00330, among others.
What is CCL2 binding?
CCL2 binding is the physical interaction between CCL2 and proteins such as CCR2, which triggers immune cell recruitment and inflammatory signaling.
How is CCL2 binding studied?
It is studied using binding assays like SPR, cell-based signaling assays, and CRISPR knockout models.
What diseases are associated with CCL2 binding?
CCL2 binding is linked to glioblastoma, esophageal cancer, MASLD, diabetic anxiety, and implantation failure.
What is the role of CCR2 in CCL2 binding?
CCR2 is the canonical signaling receptor for CCL2 and mediates monocyte chemotaxis and tumor-associated macrophage recruitment.
How does CCL2 binding affect the tumor microenvironment?
CCL2 binding promotes recruitment of immunosuppressive macrophages and supports tumor progression in glioblastoma and ESCC.
Can CCL2 binding be targeted therapeutically?
Yes, blocking CCL2 binding with small molecules or antibodies is an active therapeutic strategy for cancer and MASLD.
What is the difference between CCL2 binding and CCR2 signaling?
CCL2 binding is the physical interaction event, while CCR2 signaling refers to downstream intracellular pathways activated after binding.
What model systems are used to study CCL2 binding?
Common models include CCR2 knockout mice, CCL2-overexpressing cell lines, and bovine endometrial explants.
Conclusion
GO:0035715, chemokine (C-C motif) ligand 2 binding, is a fundamental molecular function that governs immune cell recruitment, tumor microenvironment remodeling, and metabolic inflammation. The interaction between CCL2 and its binding partners, particularly CCR2 and ACKR2, is a validated therapeutic target in cancer, liver disease, and neuroinflammation. Continued research using CRISPR-based models will refine our understanding of CCL2 binding specificity and enable the development of selective inhibitors.
References
- 1. Chen J et al.. 2024. Lnc-H19-derived protein shapes the immunosuppressive microenvironment of glioblastoma.. Cell Rep Med 5(11):101806 PMID: 39481387
- 2. Yun CS et al.. 2024. C-C motif chemokine ligand 2 regulates prostaglandin synthesis and embryo attachment of the bovine endometrium during implantation.. Cell Tissue Res 396(2):231-243 PMID: 38438567
- 3. Zhao L et al.. 2024. LINC00330/CCL2 axis-mediated ESCC TAM reprogramming affects tumor progression.. Cell Mol Biol Lett 29(1):77 PMID: 38769475
- 4. Zhu Z et al.. 2022. Chemokine (C-C motif) ligand 2-enhanced adipogenesis and angiogenesis of human adipose-derived stem cell and human umbilical vein endothelial cell co-culture system in adipose tissue engineering.. J Tissue Eng Regen Med 16(2):163-176 PMID: 34811942
- 5. Chen M et al.. 2025. Molecular mechanism of IgE-mediated FcεRI activation.. Nature 637(8045):453-460 PMID: 39442557
- 6. Pan K et al.. 2025. Neuronal CCL2 responds to hyperglycaemia and contributes to anxiety disorders in the context of diabetes.. Nat Metab 7(5):1052-1072 PMID: 40329008
- 7. Hsiang CY et al.. 2025. 2,5-Dihydroxybenzoic Acid Ameliorates Metabolic Dysfunction-Associated Steatotic Liver Disease by Targeting the CCL2-CCR2 Axis to Reduce Lipid Accumulation.. Nutrients 17(11) PMID: 40507104
- 8. Zhang J et al.. 2010. Multiple roles of chemokine (C-C motif) ligand 2 in promoting prostate cancer growth.. J Natl Cancer Inst 102(8):522-8 PMID: 20233997
