Key determinants of selective binding and activation by the monocyte chemoattractant proteins at the chemokine receptor CCR2

Huma, Zil E; Sanchez, Julie; Lim, Herman D.; Bridgford, Jessica L.; Huang, Cheng; Parker, Bradyn J.; Pazhamalil, Jiann G.; Porebski, Benjamin T.; Pfleger, Kevin D. G.; Lane, J. Robert; Canals, Meritxell; Stone, Martin J.

doi:10.1126/scisignal.aai8529

Cited by 37 publications

(42 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To avoid disrupting the folded structures of the chemokines, hydrophobic core residues were excluded from being replaced. As described in a previous study of their interactions with CCR2, all of the chimeras are well folded (20).…”

Section: Evaluation Of Site 1 and Site 2 Interactions Using Chemokinementioning

confidence: 57%

“…The form of CCL15 used in this study is the active, high-affinity form CCL15(⌬26), which has the N-terminal sequence HFAAD (52). All chemokines and chimeras were expressed and purified as described (20,46). Briefly, the N-terminal His 6 -tagged protein was expressed from BL21 (DE3) E. coli in lysogeny broth medium by induction with isopropyl 1-thio-␤-D-galactopyranoside.…”

Section: Chemokine Expression and Purificationmentioning

confidence: 99%

“…However, a number of observations point to possible deficiencies in this simple model. In particular, mutations in CS2 can influence receptor-binding affinity, suggesting that binding and activation are not simply partitioned between the two structural sites (19,20). Similarly, different chemokine agonists of the same receptor can selectively interact with different regions of a receptor (21) or can "bias" a shared receptor toward activation of different signaling pathways (22,23), suggesting distinct activated conformations of the receptor.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Evaluation and extension of the two-site, two-step model for binding and activation of the chemokine receptor CCR1

Sanchez

Huma

Lane

et al. 2019

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Interactions between secreted immune proteins called chemokines and their cognate G protein-coupled receptors regulate the trafficking of leukocytes in inflammatory responses. The two-site, two-step model describes these interactions. It involves initial binding of the chemokine N-loop/␤3 region to the receptor's N-terminal region and subsequent insertion of the chemokine N-terminal region into the transmembrane helical bundle of the receptor concurrent with receptor activation. Here, we test aspects of this model with C-C motif chemokine receptor 1 (CCR1) and several chemokine ligands. First, we compared the chemokinebinding affinities of CCR1 with those of peptides corresponding to the CCR1 N-terminal region. Relatively low affinities of the peptides and poor correlations between CCR1 and peptide affinities indicated that other regions of the receptor may contribute to binding affinity. Second, we evaluated the contributions of the two CCR1-interacting regions of the cognate chemokine ligand CCL7 (formerly monocyte chemoattractant protein-3 (MCP-3)) using chimeras between CCL7 and the non-cognate ligand CCL2 (formerly MCP-1). The results revealed that the chemokine N-terminal region contributes significantly to binding affinity but that differences in binding affinity do not completely account for differences in receptor activation. On the basis of these observations, we propose an elaboration of the two-site, two-step model-the "three-step" model-in which initial interactions of the first site result in low-affinity, nonspecific binding; rate-limiting engagement of the second site enables high-affinity, specific binding; and subsequent conformational rearrangement gives rise to receptor activation.

show abstract

Section: Evaluation Of Site 1 and Site 2 Interactions Using Chemokinementioning

confidence: 57%

Section: Chemokine Expression and Purificationmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Evaluation and extension of the two-site, two-step model for binding and activation of the chemokine receptor CCR1

Sanchez

Huma

Lane

et al. 2019

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…P672 was identified in a screen performed with the chemokine CCL8 (also known as MCP-2) and in addition binds the closely related chemokines CCL2 (MCP-1) and CCL7 (MCP-3). These three chemokines target the common receptor CCR2, and CCL7 and CCL8 are thought to be partial agonists in comparison with CCL2 ( 26 ). CCR2 is a key regulator of monocyte and macrophage migration ( 27 ), with potential roles in diverse inflammatory disease including rheumatoid arthritis and inflammatory bowel disease ( 1 ).…”

Section: Discussionmentioning

confidence: 99%

The N-terminal domain of a tick evasin is critical for chemokine binding and neutralization and confers specific binding activity to other evasins

Eaton¹,

Alenazi²,

Singh³

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

Tick chemokine-binding proteins (evasins) are an emerging class of biologicals that target multiple chemokines and show anti-inflammatory activities in preclinical disease models. Using yeast surface display, we identified a CCL8-binding evasin, P672, from the tick Rhipicephalus pulchellus. We found that P672 binds CCL8 and eight other CC-class chemokines with a Kd < 10 nm and four other CC chemokines with a Kd between 10 and 100 nm and neutralizes CCL3, CCL3L1, and CCL8 with an IC50 < 10 nm. The CC chemokine–binding profile was distinct from that of evasin 1 (EVA1), which does not bind CCL8. We also show that P672's binding activity can be markedly modulated by the location of a StrepII-His purification tag. Combining native MS and bottom-up proteomics, we further demonstrated that P672 is glycosylated and forms a 1:1 complex with CCL8, disrupting CCL8 homodimerization. Homology modeling of P672 using the crystal structure of the EVA1 and CCL3 complex as template suggested that 44 N-terminal residues of P672 form most of the contacts with CCL8. Replacing the 29 N-terminal residues of EVA1 with the 44 N-terminal residues of P672 enabled this hybrid evasin to bind and neutralize CCL8, indicating that the CCL8-binding properties of P672 reside, in part, in its N-terminal residues. This study shows that the function of certain tick evasins can be manipulated simply by adding a tag. We conclude that homology modeling helps identify regions with transportable chemokine-binding functions within evasins, which can be used to construct hybrid evasins with altered properties.

show abstract

“…Both monocyte chemoattractant protein-1 (MCP-1) and regulated upon activation normal T cell expressed and secreted (RANTES) belong to the family of chemokines, a special group of chemoattracant cytokines, which are involved in many physiological as well as pathological conditions of the female reproductive system [10]. MCP-1 is secreted by many different cell types such as monocytes, lymphocytes, and endothelial cells in response to inflammation and oxidative stress [11]. RANTES basically stimulates monocyte migration and chemotactic activity in the peritoneum [12].…”

Section: Introductionmentioning

confidence: 99%

Age at Menarche and Oxidative Stress Markers in Women with Endometriosis

Augoulea

Kindis

Karopoulou

et al. 2020

SN Compr. Clin. Med.

View full text Add to dashboard Cite

Endometriosis is one of the most frequent gynecological disorders defined as the existence and growth of endometrial tissue outside the uterine cavity. Aim of this study was to evaluate the level of six oxidative stress markers in the serum of patients with and without endometriosis. This study included 46 women who were undergoing laparoscopy. The participants were divided into two groups based on their laparoscopic results. Thirty-one women had visually and histopathologically confirmed endometriosis, whereas 15 did not. The serum level of ADMA (asymmetric dimethylarginine), MCP1 (monocyte chemoattractant protein 1), MMP2 (matrix metalloproteinases 2), MMP9 (matrix metalloproteinases 9), RANTES (regulated upon activation, normal T cell expressed and presumably secreted), and VEGF-A (vascular endothelial growth factor) as oxidative stress markers were measured and compared between the two groups. No significant difference in the ADMA, MCP1, MMP2, MMP9, RANTES, and VEGF-A serum levels was detected between the entometriosis patients and controls. The disease stage did not affect any of the markers' level either. Women with endometriosis had menarche at an earlier age compared to controls [11.7 (SD 1.09) vs 12.6 (SD 1.4), p = 0.04)]. These results suggest that there is no association between endometriosis and increased oxidative stress. The elevation and possible role of oxidative stress markers in the diagnosis and pathogenesis of endometriosis should be evaluated in more extended studies.

show abstract

Key determinants of selective binding and activation by the monocyte chemoattractant proteins at the chemokine receptor CCR2

Cited by 37 publications

References 54 publications

Evaluation and extension of the two-site, two-step model for binding and activation of the chemokine receptor CCR1

Evaluation and extension of the two-site, two-step model for binding and activation of the chemokine receptor CCR1

The N-terminal domain of a tick evasin is critical for chemokine binding and neutralization and confers specific binding activity to other evasins

Age at Menarche and Oxidative Stress Markers in Women with Endometriosis

Contact Info

Product

Resources

About