Sun D, Eccleston E D, Fallon A M
Department of Entomology, University of Minnesota, 1980 Folwell Avenue, St. Paul, Minnesota, 55108, USA.
Biochem Biophys Res Commun. 1998 Aug 19;249(2):410-5. doi: 10.1006/bbrc.1998.9150.
We have identified a 35-amino acid antibiotic cecropin secreted by an established mosquito cell line. C7-10 cells from the vector mosquito, Aedes albopictus, were incubated with heat-killed Escherichia coli, and materials secreted into the cell culture supernatant were recovered by acid precipitation. Following batch elution from Sep-Pak C18 cartridges and further purification by reverse phase high performance liquid chromatography (RP-HPLC) a predominant peak of antibacterial activity was characterized by mass spectrometry, amino acid composition analysis, and Edman degradation, yielding the sequence GGLKKLGKKLEGVGKRVFKASEKALPVAVGIKALG. Unlike other cecropins, the peptide was not amidated at the C-terminus. Aedes albopictus Cecropin A (AalCecA) is the first cecropin to be described from a mosquito vector of human disease. Consistent with the classification of mosquitoes among the Dipteran suborder Nematocera, AalCecA shares only 36% amino acid identity with cecropins from Drosophila melanogaster and other Cyclorrhaphid flies, whose mature cecropins share 80% to 100% amino acid identity.
我们从一个已建立的蚊子细胞系中鉴定出一种由35个氨基酸组成的抗菌肽天蚕素。将来自媒介蚊子白纹伊蚊的C7-10细胞与热灭活的大肠杆菌一起孵育，通过酸沉淀回收分泌到细胞培养上清液中的物质。从Sep-Pak C18柱上进行分批洗脱，并通过反相高效液相色谱（RP-HPLC）进一步纯化后，通过质谱、氨基酸组成分析和埃德曼降解对主要的抗菌活性峰进行了表征，得到的序列为GGLKKLGKKLEGVGKRVFKASEKALPVAVGIKALG。与其他天蚕素不同，该肽在C末端没有酰胺化。白纹伊蚊天蚕素A（AalCecA）是从人类疾病的蚊子媒介中描述的第一种天蚕素。与双翅目长角亚目蚊子的分类一致，AalCecA与黑腹果蝇和其他环裂亚目苍蝇的天蚕素只有36%的氨基酸同一性，而这些苍蝇成熟的天蚕素氨基酸同一性为80%至100%。
