Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Screening and Mechanism of Antagonist Peptide for CC Chemokine Receptor 1 (CCR1) Derived from Viral Macrophage Inflammatory Protein II

Sha Liu, Qing Ding, Pijin Wei, Hanxiao Sun , Xiuying Li, Guijie An, Yan Yang, Jingguang Zhou

Institute of Genomic Medicine, College of Pharmacy, Jinan University, Guangzhou 510632, China;

For correspondence:- Hanxiao Sun Email: sunhx718@163.com Tel:+8602038375022

Received: 11 February 2014 Accepted: 20 April 2014 Published: 23 May 2014

Citation: Liu S, Ding Q, Wei P, Sun H, Li X, An G, et al. Screening and Mechanism of Antagonist Peptide for CC Chemokine Receptor 1 (CCR1) Derived from Viral Macrophage Inflammatory Protein II. Trop J Pharm Res 2014; 13(5):697-704 doi: 10.4314/tjpr.v13i5.7

© 2014 The authors.
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Abstract

Purpose: To search for effective antagonist peptide of CC chemokine receptor 1 (CCR1), and evaluate the potential role and mechanism of peptide C18P derived from viral macrophage inflammatory protein II (vMIP-II).

Methods: Alignment, simulated peptide-cut, bioinformatics and protease digestion were used to screen and prepare antagonist peptide. Interactions between C18P and CCR1 were determined by radioligand binding assays and [³⁵S]GTPγS binding experiment. Chemotaxis assay was utilized to assess the potency for inducing or inhibiting peripheral blood mononuclear cells (PBMCs) migration. Ligand-induced intracellular calcium mobilization was tested by flow cytometry.

Results: A peptide containing 18 amino acids (C18P) was screened. C18P bound to CCR1 with a Kd of 5.7 ng/ml and displaced ¹²⁵I-labeled MIP-1α and ¹²⁵I-labeled RANTES on human CCR1-transfected HEK293 cells (IC₅₀= 11.2 and 9.6 ng/ml, respectively) in radioligand binding studies. C18P lacked intrinsic agonist activity but strongly inhibited HCC-1 (100 nM) induced [³⁵S]GTPγS binding (IC₅₀= 3.7 ug/ml), chemotaxis induced by HCC-1, MIP-1α and RANTES (IC₅₀= 23, 25 and 13.1 ng/ml, respectively), and intracellular calcium mobilization.

Conclusion: These results demonstrate that bioinformatics and protease digestion are feasible to screen and prepare C18P, and that C18P is a novel and specific small molecule peptide antagonist of CCR1 with therapeutic potential for preventing cell migration.

Keywords: CC Chemokine receptor 1; Simulated peptide-cut; Antagonist peptide; Viral macrophage inflammatory protein II; Bioinformatics; Protease digestion; HEK2