Relationship between structures and biological activities of mycoplasmal diacylated lipopeptides and their recognition by toll-like receptors 2 and 6
Relationship between structures and biological activities of mycoplasmal diacylated lipopeptides and their recognition by toll-like receptors 2 and 6
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DOI：
10.1128/iai.72.3.1657-1665.2004
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发表时间：
2004-03-01
影响因子：
3.1
通讯作者：
Shibata, KI
中科院分区：
文献类型：
--
作者：
Okusawa, T;Fujita, M;Shibata, KI
脂肽FSL-1 [S-（2，3-双棕榈酰氧基丙基）-Cys-Gly-Asp-Pro-Lys-His-Pro-Lys-Ser-Phe，Pam（2）CG DPKHPKSF]是基于能够活化正常人牙龈成纤维细胞以诱导ICAM-1的细胞表面表达的唾液支原体脂蛋白的N-末端结构合成的，显示出诱导单核细胞趋化蛋白1、白细胞介素-6（IL-6）、和IL-8。FSL-1还激活巨噬细胞产生肿瘤坏死因子α，就像支原体发酵剂衍生的脂肽MALP-2（Pam（2）CGNNDESNISFKEK）（一种有效的巨噬细胞激活脂肽）一样。FSL-1的活性水平高于MALP-2。该结果表明，肽部分的氨基酸序列的差异影响活性，因为前者的氨基酸序列以外的骨架结构与后者的相同。为了确定FSL-1活性的最低结构要求，检查二酰基甘油基Cys和肽部分的该活性。这两个部分都没有显示活性。从Phe到Arg的单个氨基酸取代和从棕榈酸到硬脂酸的脂肪酸取代大大降低了活性。在测量FSL-1对用Toll样受体2和6以及NF-κ B依赖性荧光素酶报告质粒转染的人胚肾293细胞的NF-κ B报告活性中获得了类似的结果。这些结果表明，FSL-1的二酰基甘油和肽部分对于生物活性的表达和Toll样受体2和6的识别都是必不可少的，并且Toll样受体2和6对FSL-1的识别似乎是疏水性的。
The lipopeptide FSL-1 [S-(2,3-bispalmitoyloxypropyl)-Cys-Gly-Asp-Pro-Lys-His-Pro-Lys-Ser-Phe, Pam(2)CG DPKHPKSF] synthesized on the basis of the N-terminal structure of a Mycoplasma salivarium lipoprotein capable of activating normal human gingival fibroblasts to induce the cell surface expression of ICAM-1 revealed an activity to induce production of monocyte chemoattractant protein 1, interleukin-6 (IL-6), and IL-8. FSL-1 also activated macrophages to produce tumor necrosis factor alpha as the Mycoplasmafermentans-derived lipopeptide MALP-2 (Pam(2)CGNNDESNISFKEK), a potent macrophage-activating lipopeptide, did. The level of the activity of FSL-1 was higher than that of MALP-2. This result suggests that the difference in the amino acid sequence of the peptide portion affects the activity because the framework structure other than the amino acid sequence of the former is the same as that of the latter. To determine minimal structural requirements for the activity of FSL-1, the diacylglyceryl Cys and the peptide portions were examined for this activity. Both portions did not reveal the activity. A single amino acid substitution from Phe to Arg and a fatty acid substitution from palmitic acid to stearic acid drastically reduced the activity. Similar results were obtained in measuring the NF-kappaB reporter activity of FSL-1 to human embryonic kidney 293 cells transfected with Toll-like receptor 2 and 6, together with a NF-kappaB-dependent luciferase reporter plasmid. These results suggest that both the diacylglyceryl and the peptide portions of FSL-1 are indispensable for the expression of biological activities and for the recognition by Toll-like receptors 2 and 6 and that the recognition of FSL-1 by Toll-like receptors 2 and 6 appears to be hydrophobic.