Human acetyl-CoA carboxylase 2 expressed in silkworm Bombyx mori exhibits posttranslational biotinylation and phosphorylation

In Wook Hwang, Yu Makishima, Tatsuya Kato, Sungjo Park, Andre Terzic, Enoch Y. Park

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)8201-8209
Number of pages9
JournalApplied Microbiology and Biotechnology
Volume98
Issue number19
DOIs
StatePublished - Oct 2014

Keywords

  • Bombyx mori nucleopolyhedrovirus
  • Human acetyl-CoA carboxylase 2 (hACC2)
  • Lipid metabolism
  • Phosphorylation/dephosphorylation
  • Silkworm

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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