title: Human acetyl-CoA carboxylase 2 expressed in silkworm Bombyx mori exhibits posttranslational biotinylation and phosphorylation.
abstract: Biotin-dependent human acetyl-CoA carboxylases (ACCs) are integral in homeostatic lipid metabolism. By securing posttranslational biotinylation, ACCs perform coordinated catalytic functions allosterically regulated by phosphorylation/dephosphorylation and citrate. The production of authentic recombinant ACCs is heeded to provide a reliable tool for molecular studies and drug discovery. Here, we examined whether the human ACC2 (hACC2), an isoform of ACC produced using the silkworm BmNPV bacmid system, is equipped with proper posttranslational modifications to carry out catalytic functions as the silkworm harbors an inherent posttranslational modification machinery. Purified hACC2 possessed genuine biotinylation capacity probed by biotin-specific streptavidin and biotin antibodies. In addition, phosphorylated hACC2 displayed limited catalytic activity whereas dephosphorylated hACC2 revealed an enhanced enzymatic activity. Moreover, hACC2 polymerization, analyzed by native page gel analysis and atomic force microscopy imaging, was allosterically regulated by citrate and the phosphorylation/dephosphorylation modulated citrate-induced hACC2 polymerization process. Thus, the silkworm BmNPV bacmid system provides a reliable eukaryotic protein production platform for structural and functional analysis and therapeutic drug discovery applications implementing suitable posttranslational biotinylation and phosphorylation.
authors.affiliation: Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan.
authors.forename: In-Wook
authors.initials: IW
authors.lastname: Hwang
authors.forename: Yu
authors.initials: Y
authors.lastname: Makishima
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Kato
authors.forename: Sungjo
authors.initials: S
authors.lastname: Park
authors.forename: Andre
authors.initials: A
authors.lastname: Terzic
authors.forename: Enoch Y
authors.initials: EY
authors.lastname: Park
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D019921:Biotinylation; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational
chemical_list: C520105:ACACB protein, human; D000103:Acetyl-CoA Carboxylase
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