2DZC

Crystal Structure Of Biotin Protein Ligase From Pyrococcus Horikoshii, Mutation R48A

2DZC の概要

• エントリーDOI: 10.2210/pdb2dzc/pdb
• 関連するPDBエントリー: 2DXU
• 分子名称: biotin--[acetyl-CoA-carboxylase] ligase (2 entities in total)
• 機能のキーワード: biotin biosynthesis, dimer, structural genomics, nppsfa, national project on protein structural and functional analyses, riken structural genomics/proteomics initiative, rsgi, ligase
• 由来する生物種: Pyrococcus horikoshii
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 52034.82

構造登録者

Bagautdinov, B.,Taketa, M.,Matsuura, Y.,Kunishima, N.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (登録日: 2006-09-27, 公開日: 2007-03-27, 最終更新日: 2023-10-25)

主引用文献

Bagautdinov, B.,Matsuura, Y.,Bagautdinova, S.,Kunishima, N.
Protein biotinylation visualized by a complex structure of biotin protein ligase with a substrate
J.Biol.Chem., 283:14739-14750, 2008

PubMed Abstract: Biotin protein ligase (BPL) catalyzes the biotinylation of the biotin carboxyl carrier protein (BCCP) only at a special lysine residue. Here we report the first structure of BPL.BCCP complex crystals, which are prepared using two BPL mutants: R48A and R48A/K111A. From a detailed structural characterization, it is likely that the mutants retain functionality as enzymes but have a reduced activity to produce the reaction intermediate biotinyl-5'-AMP. The observed biotin and partly disordered ATP in the mutant structures may act as a non-reactive analog of the substrates or biotinyl-5'-AMP, thereby providing the complex crystals. The four crystallographically independent BPL.BCCP complexes obtained can be classified structurally into three groups: the formation stages 1 and 2 with apo-BCCP and the product stage with biotinylated holo-BCCP. Residues responsible for the complex formation as well as for the biotinylation reaction have been identified. The C-terminal domain of BPL shows especially large conformational changes to accommodate BCCP, suggesting its functional importance. The formation stage 1 complex shows the closest distance between the carboxyl carbon of biotin and the special lysine of BCCP, suggesting its relevance to the unobserved reaction stage. Interestingly, bound ATP and biotin are also seen in the product stage, indicating that the substrates may be recruited into the product stage complex before the release of holo-BCCP, probably for the next reaction cycle. The existence of formation and product stages before and after the reaction stage would be favorable to ensure both the reaction efficiency and the extreme substrate specificity of the biotinylation reaction.

PubMed: 18372281
DOI: 10.1074/jbc.M709116200

実験手法

X-RAY DIFFRACTION (1.45 Å)