4WL2

Structure of penicillin V acylase from Pectobacterium atrosepticum

4WL2 の概要

• エントリーDOI: 10.2210/pdb4wl2/pdb
• 分子名称: Putative exported choloylglycine hydrolase (2 entities in total)
• 機能のキーワード: ntn hydrolase, choloylglycine, penicillin v, periplasmic, hydrolase
• 由来する生物種: Pectobacterium atrosepticum SCRI1043
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 315802.31

構造登録者

Ramasamy, S.,Avinash, V.S.,Pundle, A.V.,Suresh, C.G. (登録日: 2014-10-06, 公開日: 2015-10-07, 最終更新日: 2024-10-16)

主引用文献

Avinash, V.S.,Panigrahi, P.,Chand, D.,Pundle, A.,Suresh, C.G.,Ramasamy, S.
Structural analysis of a penicillin V acylase from Pectobacterium atrosepticum confirms the importance of two Trp residues for activity and specificity
J.Struct.Biol., 193:85-94, 2016

PubMed Abstract: Penicillin V acylases (PVA) catalyze the deacylation of the beta-lactam antibiotic phenoxymethylpenicillin (Pen V). They are members of the Ntn hydrolase family and possess an N-terminal cysteine as the main catalytic nucleophile residue. They form the evolutionarily related cholylglycine hydrolase (CGH) group which includes bile salt hydrolases (BSH) responsible for bile deconjugation. Even though a few PVA and BSH structures have been reported, no structure of a functional PVA from Gram-negative bacteria is available. Here, we report the crystal structure of a highly active PVA from Gram-negative Pectobacterium atrosepticum (PaPVA) at 2.5Å resolution. Structural comparison with PVAs from Gram-positive bacteria revealed that PaPVA had a distinctive tetrameric structure and active site organization. In addition, mutagenesis of key active site residues and biochemical characterization of the resultant variants elucidated the role of these residues in substrate binding and catalysis. The importance of residue Trp23 and Trp87 side chains in binding and correct positioning of Pen V by PVAs was confirmed using mutagenesis and substrate docking with a 15ns molecular dynamics simulation. These results establish the unique nature of Gram-negative CGHs and necessitate further research about their substrate spectrum.

PubMed: 26707624
DOI: 10.1016/j.jsb.2015.12.008

実験手法

X-RAY DIFFRACTION (2.5 Å)