Degradation of Manduca sexta allatostatin and allatotropin by proteases associated with the foregut of Lacanobia oleracea larvae

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• 作者：Audsley ; Neil ; Weaver ; Robert J. ; Edwards ; John P.
• 关键词：Neuropeptide ; Metabolism ; MALDI-TOF ; Mass spectrometry ; Insect ; Protease inhibitor
• 刊名：Peptides
• 出版年：2002
• 出版时间：November, 2002
• 年：2002
• 卷：23
• 期：11
• 页码：2015-2023
• 全文大小：132 K

文摘

The degradation of synthetic Manduca sexta allatostatin (Manse-AS) and allatotropin (Manse-AT), by enzymes of the foregut of larvae of the tomato moth, Lacanobia oleracea was investigated using reversed-phase high performance liquid chromatography (RP-HPLC) together with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Edman sequencing. Metabolism of 1nmol Manse-AS by foregut extract (1μg protein) was rapid, t_1/2es/glyphs/BQ4.GIF>5min, with two major products produced. Mass spectrometry of HPLC fractions identified cleavage products Manse-AS-(4-15) and Manse-AS-(6-15), which indicates enzymatic cleavage at the C-terminal side of arginine residues (R³ and R⁵). This degradation of Manse-AS could be inhibited by up to 80 % by the serine protease inhibitor aprotinin, but not PMSF, pepstatin, E64, EDTA, or 1,10-phenanthroline.