scholarly journals Site-directed mutagenesis of monocyte chemoattractant protein-1 identifies two regions of the polypeptide essential for biological activity

1996 ◽  
Vol 313 (2) ◽  
pp. 633-640 ◽  
Author(s):  
Clifford J. BEALL ◽  
Sangeeta MAHAJAN ◽  
Donald E. KUHN ◽  
Pappachan E. KOLATTUKUDY
Keyword(s):  
Biological Activity ◽  
Inflammatory Diseases ◽  
Expression System ◽  
Point Mutations ◽  
Site Directed Mutagenesis ◽  
Monocyte Chemoattractant Protein 1 ◽  
Monocyte Chemoattractant ◽  
Monocyte Chemoattractant Protein ◽  
Monocyte Migration ◽  
Important Region

Monocyte chemoattractant protein-1 (MCP-1) mediates monocyte migration into tissues in inflammatory diseases and atherosclerosis. We have investigated structure-activity relationships for human MCP-1. Mutations were introduced based upon differences between MCP-1 and the structurally related but functionally distinct molecule interleukin-8 (IL-8). Mutant proteins produced using the baculovirus/insect cell expression system were purified and their ability to stimulate monocyte chemotaxis and elevation of intracellular calcium in THP-1 monocytic leukaemia cells was measured. Two regions in MCP-1 were identified as important for its biological activity. One region consists of the sequence Thr-Cys-Cys-Tyr (amino acids 10-13). Point mutations of Thr-10 to Arg and Tyr-13 to Ile greatly lowered MCP-1 activity. The second functionally important region is formed by Ser-34 and Lys-35. Insertion of a Pro between these two residues, or their substitution by the sequence Gly-Pro-His, caused nearly complete loss of MCP-1 activity. Competition binding experiments showed that the mutations that affected activity also lowered the ability to compete with wild-type MCP-1 for receptors on THP-1 cells. Point mutations at positions 8, 15, 30, 37, 38 and 68 had little effect on MCP-1 activity. The important regions that we have identified in MCP-1 correspond with previously identified functionally important regions of IL-8, suggesting that the two molecules bind to their respective receptors by similar contacts.

scholarly journals Association between monocyte chemoattractant protein-1 and blood pressure in smokers

2017 ◽  
Vol 46 (3) ◽  
pp. 965-974 ◽  
Author(s):  
Maki Komiyama ◽  
Rieko Takanabe ◽  
Koh Ono ◽  
Sayaka Shimada ◽  
Hiromichi Wada ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Systolic Blood Pressure ◽  
Atherosclerotic Lesion ◽  
The Body ◽  
Enzyme Linked Immunosorbent Assay ◽  
Monocyte Chemoattractant Protein 1 ◽  
Monocyte Chemoattractant ◽  
Monocyte Chemoattractant Protein ◽  
Monocyte Migration ◽  
Smoking Duration

Objective The expression level of monocyte chemoattractant protein-1 (MCP-1) is increased in atherosclerotic regions, inducing monocyte migration to the blood vessel wall. Although the serum MCP-1 concentration is higher in patients with than without cardiovascular disease, the precise correlations between the serum MCP-1 concentration and factors associated with smoking and atherosclerosis are unknown. Methods The serum MCP-1 concentration was measured using an enzyme-linked immunosorbent assay in 207 consecutive smokers who visited our smoking cessation clinic. Results Sex-adjusted analysis of smokers revealed that the MCP-1 concentration was positively correlated with age ( β = 0.311), smoking duration ( β = 0.342), systolic blood pressure ( β = 0.225), and diastolic blood pressure ( β = 0.137) but not with the body mass index. Multivariate regression analysis showed that smoking duration and systolic blood pressure were independent determinants of the MCP-1 concentration. Conclusions The MCP-1 concentration was positively correlated with blood pressure among smokers. Long-term smokers with high blood pressure may be more susceptible to plaque rupture at atherosclerotic lesion sites.

Homocysteine induces monocyte chemoattractant protein-1 expression by activating NF-κB in THP-1 macrophages

2001 ◽  
Vol 280 (6) ◽  
pp. H2840-H2847 ◽  
Author(s):  
Guoping Wang ◽  
Yaw L. Siow ◽  
Karmin O
Keyword(s):  
Underlying Mechanism ◽  
Inhibitory Protein ◽  
Monocytic Cell ◽  
Monocyte Chemoattractant Protein 1 ◽  
Arterial Walls ◽  
Monocyte Chemoattractant ◽  
Monocyte Chemoattractant Protein ◽  
Monocyte Migration ◽  
Human Monocytic Cell ◽  
Monocyte Chemotaxis

Homocysteinemia is an independent risk factor for cardiovascular disorders. The recruitment of monocytes is an important event in atherogenesis. Monocyte chemoattractant protein-1 (MCP-1) is a potent chemokine that stimulates monocyte migration into the intima of arterial walls. The objective of the present study was to investigate the effect of homocysteine on MCP-1 expression in macrophages and the underlying mechanism of such effect. Human monocytic cell (THP-1)-derived macrophages were incubated with homocysteine. By nuclease protection assay and ELISA, homocysteine (0.05–0.2 mM) was shown to significantly enhance the expression of MCP-1 mRNA (up to 2.6-fold) and protein (up to 4.8-fold) in these cells. Homocysteine-induced MCP-1 expression resulted in increased monocyte chemotaxis. The increase in MCP-1 expression was associated with activation of nuclear factor (NF)-κB due to increased phosphorylation of the inhibitory protein (IκB-α) as well as reduced expression of IκB-α mRNA in homocysteine-treated cells. In conclusion, our results demonstrate that homocysteine, at pathological concentration, stimulates MCP-1 expression in THP-1 macrophages via NF-κB activation.

scholarly journals Chondroitin Sulfate Inhibits Monocyte Chemoattractant Protein-1 Release From 3T3-L1 Adipocytes: A New Treatment Opportunity for Obesity-Related Inflammation?

2017 ◽  
Vol 12 ◽  
pp. 117727191772696 ◽  
Author(s):  
Thomas V Stabler ◽  
Eulàlia Montell ◽  
Josep Vergés ◽  
Janet L Huebner ◽  
Virginia Byers Kraus
Keyword(s):  
Adipose Tissue ◽  
Chondroitin Sulfate ◽  
Dependent Manner ◽  
Monocyte Chemoattractant Protein 1 ◽  
Monocyte Chemoattractant ◽  
Monocyte Chemoattractant Protein ◽  
Monocyte Migration ◽  
Inflammatory Stimuli ◽  
New Treatment

Monocyte chemoattractant protein-1 (MCP-1) overproduction from inflamed adipose tissue is a major contributor to obesity-related metabolic syndromes. 3T3-L1 embryonic fibroblasts were cultured and differentiated into adipocytes using an established protocol. Adipocytes were treated with lipopolysaccharide (LPS) to induce inflammation and thus MCP-1 release. At the same time, varying concentrations of chondroitin sulfate (CS) were added in a physiologically relevant range (10-200 µg/mL) to determine its impact on MCP-1 release. Chondroitin sulfate, a natural glycosaminoglycan of connective tissue including the cartilage extracellular matrix, was chosen on the basis of our previous studies demonstrating its anti-inflammatory effect on macrophages. Because the main action of MCP-1 is to induce monocyte migration, cultured THP-1 monocytes were used to test whether CS at the highest physiologically relevant concentration could inhibit cell migration induced by human recombinant MCP-1. Chondroitin sulfate (100-200 µg/mL) inhibited MCP-1 release from inflamed adipocytes in a dose-dependent manner ( P < .01, 95% confidence interval [CI]: −5.89 to −3.858 at 100 µg/mL and P < .001, 95% CI: −6.028 to −3.996 at 200 µg/mL) but had no effect on MCP-1–driven chemotaxis of THP-1 monocytes. In summary, CS could be expected to reduce macrophage infiltration into adipose tissue by reduction in adipocyte expression and release of MCP-1 and as such might reduce adipose tissue inflammation in response to pro-inflammatory stimuli such as LPS, now increasingly recognized to be relevant in vivo.

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