8CC5

Vibrio cholerae GbpA (LPMO domain)

8CC5 の概要

• エントリーDOI: 10.2210/pdb8cc5/pdb
• 分子名称: GlcNAc-binding protein A, ACETATE ION, COPPER (II) ION, ... (6 entities in total)
• 機能のキーワード: adhesin, lpmo, redox enzyme, colonization factor, cell adhesion
• 由来する生物種: Vibrio cholerae O1 biovar El Tor
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 40433.42

構造登録者

Montserrat-Canals, M.,Sorensen, H.V.,Cordara, G.,Krengel, U. (登録日: 2023-01-26, 公開日: 2023-08-30)

主引用文献

Sorensen, H.V.,Montserrat-Canals, M.,Loose, J.S.M.,Fisher, S.Z.,Moulin, M.,Blakeley, M.P.,Cordara, G.,Bjerregaard-Andersen, K.,Krengel, U.
Perdeuterated GbpA Enables Neutron Scattering Experiments of a Lytic Polysaccharide Monooxygenase.
Acs Omega, 8:29101-29112, 2023

PubMed Abstract: Lytic polysaccharide monooxygenases (LPMOs) are surface-active redox enzymes that catalyze the degradation of recalcitrant polysaccharides, making them important tools for energy production from renewable sources. In addition, LPMOs are important virulence factors for fungi, bacteria, and viruses. However, many knowledge gaps still exist regarding their catalytic mechanism and interaction with their insoluble, crystalline substrates. Moreover, conventional structural biology techniques, such as X-ray crystallography, usually do not reveal the protonation state of catalytically important residues. In contrast, neutron crystallography is highly suited to obtain this information, albeit with significant sample volume requirements and challenges associated with hydrogen's large incoherent scattering signal. We set out to demonstrate the feasibility of neutron-based techniques for LPMOs using -acetylglucosamine-binding protein A (GbpA) from as a target. GbpA is a multifunctional protein that is secreted by the bacteria to colonize and degrade chitin. We developed an efficient deuteration protocol, which yields >10 mg of pure 97% deuterated protein per liter expression media, which was scaled up further at international facilities. The deuterated protein retains its catalytic activity and structure, as demonstrated by small-angle X-ray and neutron scattering studies of full-length GbpA and X-ray crystal structures of its LPMO domain (to 1.1 Å resolution), setting the stage for neutron scattering experiments with its substrate chitin.

PubMed: 37599915
DOI: 10.1021/acsomega.3c02168

実験手法

X-RAY DIFFRACTION (1.62 Å)