Proteomic identification, cDNA cloning and enzymatic activity of glutathione S

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• 作者：Kristen E. Whalen ; Dexter Morin ; Ching Yu Lin ; Ronald S. Tjeerdema ; Jared V. Goldstone ; Mark E.Hahn
• 关键词：Cyphoma gibbosum ; Glutathione S-transferase ; Gorgonian ; Natural product ; Chemical ecology ; Allelochemical ; Proteomic
• 刊名：Archives of Biochemistry and Biophysics
• 出版年：2008
• 出版时间：1 October 2008
• 年：2008
• 卷：478
• 期：1
• 页码：7-17
• 全文大小：513 K

文摘

Glutathione S-transferases (GST) were characterized from the digestive gland of Cyphoma gibbosum (Mollusca; Gastropoda), to investigate the possible role of these detoxification enzymes in conferring resistance to allelochemicals present in its gorgonian coral diet. We identified the collection of expressed cytosolic Cyphoma GST classes using a proteomic approach involving affinity chromatography, HPLC and nano-spray liquid chromatography–tandem mass spectrometry (LC–MS/MS). Two major GST subunits were identified as putative mu-class GSTs; while one minor GST subunit was identified as a putative theta-class GST, apparently the first theta-class GST identified from a mollusc. Two Cyphoma GST cDNAs (CgGSTM1 and CgGSTM2) were isolated by RT-PCR using primers derived from peptide sequences. Phylogenetic analyses established both cDNAs as mu-class GSTs and revealed a mollusc-specific subclass of the GST-mu clade. These results provide new insights into metazoan GST diversity and the biochemical mechanisms used by marine organisms to cope with their chemically defended prey.