UM
Interactions of peptides from secreted human CKLF1 and the N-terminal extracellular tail of CCR4 analyzed by CZE
Sun Z.1; Ling X.1; Zhang Y.2; Tian L.2; Wang Y.2
2009-06-03
Source PublicationChromatographia
ISSN00095893 16121112
Volume70Issue:1-2Pages:287-292
AbstractHuman 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 ± 0.86) × 10 M. The binding constant of the interactions between CKLF1-C27/CKLF1-C19 and ML40 was calculated as (2.96 ± 0.59) × 10 M and (1.39 ± 0.38) × 10 M 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. © 2009 Vieweg+Teubner | GWV Fachverlage GmbH.
KeywordCapillary zone electrophoresis CKLF1-C27, CKLF1-C19, CKLF1-C19km, CKLF1-C19pm and ML40 Peptides
DOI10.1365/s10337-009-1151-7
URLView the original
Language英語
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Cited Times [WOS]:3   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionUniversity of Macau
Affiliation1.Peking University
2.Peking University Health Science Center
Recommended Citation
GB/T 7714
Sun Z.,Ling X.,Zhang Y.,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 Z.,Ling X.,Zhang Y.,Tian L.,&Wang Y..(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 Z.,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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