Lackmann M, Rajasekariah P, Iismaa S E, Jones G, Cornish C J, Hu S, Simpson R J, Moritz R L, Geczy C L
Heart Research Institute, Camperdown, NSW, Australia.
J Immunol. 1993 Apr 1;150(7):2981-91.
We previously reported the purification and partial amino acid sequence of a novel murine cytokine designated CP-10, which has chemotactic activity for murine polymorphonuclear cells (PMN) and macrophages. The complete cDNA encoding an 88-amino acid polypeptide has been isolated and the sequence is presented here. Transient transfection of CP-10 cDNA into CV-1 cells confirmed the chemotactic activity of rCP-10 for murine PMN. CP-10 has sequence homology with members of the S100 family of Ca(2+)-binding proteins with pronounced amino acid sequence similarities within the putative N- and C-terminal Ca(2+)-binding sites, but differences within their connecting hinge and C-terminal regions. We have confirmed the hypothesis of Kligman and Hilt that functional specificity of individual members of the S100 protein family may reside in the hinge region. A synthetic peptide corresponding to the hinge region of CP-10 (CP-10(42-55) was compared with native CP-10 in chemotaxis and skin test assays. Native CP-10 had potent activity for phagocytic cells, but not lymphocytes, in vitro (optimal activity, 10(-11) to 10(-13) M) and elicited a sustained recruitment of neutrophils and mononuclear cells over 24 h in vivo. The hinge-region peptide had strong chemotactic activity for murine phagocytic cells (optimal activity, 10(-10) - 10(-11) M) but elicited only a transient infiltration of neutrophils over 4 to 8 h after intradermal injection. Results indicate that although the hinge region contributes significantly to the functional specificity of the S100 protein CP-10, sustained cellular recruitment typical of a delayed type hypersensitivity response is apparently dependent on the structural integrity of the protein.
我们之前报道了一种名为CP-10的新型小鼠细胞因子的纯化及部分氨基酸序列，该细胞因子对小鼠多形核细胞（PMN）和巨噬细胞具有趋化活性。现已分离出编码88个氨基酸多肽的完整cDNA，并在此展示其序列。将CP-10 cDNA瞬时转染到CV-1细胞中，证实了重组CP-10对小鼠PMN具有趋化活性。CP-10与Ca(2+)结合蛋白S100家族成员具有序列同源性，在假定的N端和C端Ca(2+)结合位点内有明显的氨基酸序列相似性，但在其连接铰链区和C端区域存在差异。我们证实了Kligman和Hilt的假设，即S100蛋白家族单个成员的功能特异性可能存在于铰链区。在趋化性和皮肤试验中，将与CP-10铰链区对应的合成肽（CP-10(42-55)）与天然CP-10进行了比较。天然CP-10在体外对吞噬细胞具有强大活性，但对淋巴细胞无活性（最佳活性为10(-11)至10(-13) M），并在体内24小时内引起中性粒细胞和单核细胞的持续募集。铰链区肽对小鼠吞噬细胞具有强烈的趋化活性（最佳活性为10(-10) - 10(-11) M），但皮内注射后仅在4至8小时内引起中性粒细胞的短暂浸润。结果表明，尽管铰链区对S100蛋白CP-10的功能特异性有显著贡献，但迟发型超敏反应典型的持续细胞募集显然依赖于该蛋白的结构完整性。
