title: Structural and functional characterization of S-adenosylmethionine (SAM) synthetase from Pichia ciferrii.
abstract: S-adenosylmethionine synthetase (SAM-s) catalyzes the synthesis of S-adenosylmethionine (SAM), which is essential for methylation, transcription, proliferation, and production of secondary metabolites. Here SAM-s from Pichia ciferrii were selectively cloned using RNA CapFishing and rapid amplification of cDNA ends (RACE). The putative full-length cDNA of SAM-s encoded a 383 amino acid protein (42.6 kDa), which has highly conserved metal binding sites, a phosphate-binding site, and functionally important motifs. The corresponding enzyme was over-expressed in a heterologous host of Pichia pastoris, and then purified to a homogenous form. Enzyme kinetics, immunoblotting, circular dichroism (CD), high performance liquid chromatography (HPLC), and molecular modeling were conducted to characterize the SAM-s from P. ciferrii. Structural and functional studies of SAM-s will provide important insights for industrial applications.
authors: Department of Molecular Science and Technology, Graduate School of Interdisciplinary Program, Ajou University, Suwon, 443-749, Korea.
authors: Sangyoung
authors: S
authors: Yoon
authors: Wonkyu
authors: W
authors: Lee
authors: Minsoo
authors: M
authors: Kim
authors: T Doohun
authors: TD
authors: Kim
authors: Yeonwoo
authors: Y
authors: Ryu
mesh_terms: D000595:Amino Acid Sequence; D004789:Enzyme Activation; D004795:Enzyme Stability; D008716:Methionine Adenosyltransferase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010843:Pichia / Q000145:classification / Q000201:enzymology*; D011487:Protein Conformation; D012436:S-Adenosylmethionine / Q000737:chemistry* / Q000648:ultrastructure*; D013045:Species Specificity
chemical_list: D012436:S-Adenosylmethionine; D008716:Methionine Adenosyltransferase