Proteomic identification, cDNA cloning and enzymatic activity of glutathione S-transferases from the generalist marine gastropod, Cyphoma gibbosum.
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.
Whalen
Kristen E
KE
Biology Department, Woods Hole Oceanographic Institution, MS 32, 45 Water Street, Woods Hole, MA 02543, USA. kwhalen@whoi.edu
Morin
Dexter
D
Lin
Ching Yu
CY
Tjeerdema
Ronald S
RS
Goldstone
Jared V
JV
Hahn
Mark E
ME
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D049851:Gastropoda; D005982:Glutathione Transferase / Q000378:metabolism*; D013058:Mass Spectrometry; D008954:Models, Biological; D008969:Molecular Sequence Data; D010802:Phylogeny; D040901:Proteomics / Q000379:methods*; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D005982:Glutathione Transferase