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学科主题药学
Interactions of Peptides from Secreted Human CKLF1 and the N-Terminal Extracellular Tail of CCR4 Analyzed by CZE
Sun, Zhe1; Ling, Xiaomei1; Zhang, Yingmei2,3; Tian, Linjie2; Wang, Ying2,3
关键词Capillary Zone Electrophoresis Peptides Cklf1-c27 Cklf1-c19 Cklf1-c19km Cklf1-c19pm And Ml40
刊名CHROMATOGRAPHIA
2009-07-01
DOI10.1365/s10337-009-1151-7
70期:1-2页:287-292
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemical Research Methods ; Chemistry, Analytical
资助者National High Technology Research and Development Program of China ; National Natural Science Foundation of China ; National High Technology Research and Development Program of China ; National Natural Science Foundation of China
研究领域[WOS]Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]CHEMOKINE-LIKE FACTOR-1 ; CAPILLARY-ELECTROPHORESIS ; FUNCTIONAL LIGAND ; MCP-1 RECEPTOR ; BINDING ; IDENTIFICATION ; FLUORESCENCE ; FRACTALKINE ; MECHANISM ; FRAGMENT
英文摘要

Human chemokine-like factor 1 (CKLF1) exhibits chemotactic effects on leukocytes. A previous study demonstrated that CKLF1 is a functional ligand for human CC chemokine receptor 4 (CCR4). The N-terminal amino acid sequencing of secreted CKLF1 protein showed that it contains at least two peptides, CKLF1-C27 and CKLF1-C19, which have functional activation via CCR4. To quantitatively evaluate the interaction of CKLF1-C27 or CKLF1-C19 with CCR4, the N-terminal extracellular tail of CCR4 (ML40), CKLF1-C27 and CKLF1-C19 were chemically synthesized and analyzed by capillary zone electrophoresis. Both qualitative and quantitative characterizations of the peptide-peptide binding were determined. We used the macrophage-derived chemokine (MDC) as the positive control and its binding constant was (4.99 +/- A 0.86) x 10(4) M-1. The binding constant of the interactions between CKLF1-C27/CKLF1-C19 and ML40 was calculated as (2.96 +/- A 0.59) x 10(4) M-1 and (1.39 +/- A 0.38) x 10(4) M-1 by the Scatchard analysis. This result proved that CKLF1-C27 had a greater potent affinity with ML40 than CKLF1-C19 because of the excess eight amino acids. To understand the molecular basis for the interaction, a mutagenesis study of CKLF1-C19 was undertaken. CKLF1-C19pm and CKLF1-C19km were synthesized and their interactions with ML40 were analyzed by CZE. We found that the substitution of Lys or Pro to Ala within the residues of CKLF1-C19 strongly abolished or decreased its interaction with ML40, suggesting that the Lys residues of CKLF1-C19 play the important role for the interaction and the Pro residues of CKLF1-C19 affect its affinity. All the experimental results show that the reported method by CZE for the determination of the interactions of CKLF1 peptides and the N-terminal extracellular tail of CCR4 is powerful.

语种英语
所属项目编号2006AA02A305 ; 30671907 ; 30872292 ; 90813025
资助者National High Technology Research and Development Program of China ; National Natural Science Foundation of China ; National High Technology Research and Development Program of China ; National Natural Science Foundation of China
WOS记录号WOS:000268055600041
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/57002
Collection北京大学药学院_化学生物学系
作者单位1.Peking Univ, Dept Pharmaceut Anal, Sch Pharmaceut Sci, Beijing 100083, Peoples R China
2.Peking Univ, Lab Med Immunol, Sch Basic Med Sci, Hlth Sci Ctr, Beijing 100083, Peoples R China
3.Peking Univ, Ctr Human Dis Genom, Beijing 100083, Peoples R China
Recommended Citation
GB/T 7714
Sun, Zhe,Ling, Xiaomei,Zhang, Yingmei,et al. Interactions of Peptides from Secreted Human CKLF1 and the N-Terminal Extracellular Tail of CCR4 Analyzed by CZE[J]. CHROMATOGRAPHIA,2009,70(1-2):287-292.
APA Sun, Zhe,Ling, Xiaomei,Zhang, Yingmei,Tian, Linjie,&Wang, Ying.(2009).Interactions of Peptides from Secreted Human CKLF1 and the N-Terminal Extracellular Tail of CCR4 Analyzed by CZE.CHROMATOGRAPHIA,70(1-2),287-292.
MLA Sun, Zhe,et al."Interactions of Peptides from Secreted Human CKLF1 and the N-Terminal Extracellular Tail of CCR4 Analyzed by CZE".CHROMATOGRAPHIA 70.1-2(2009):287-292.
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