5OVM

Solution structure of lipase binding domain LID1 of foldase from Pseudomonas aeruginosa

5OVM の概要

• エントリーDOI: 10.2210/pdb5ovm/pdb
• NMR情報: BMRB: 34175
• 分子名称: Lipase chaperone (1 entity in total)
• 機能のキーワード: lipase a, lipase interaction domain 1, chaperon, pseudomonas aeruginosa, chaperone
• 由来する生物種: Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 10108.32

構造登録者

Viegas, A.,Jaeger, K.-E.,Etzkorn, M.,Gohlke, H.,Verma, N.,Dollinger, P.,Kovacic, F. (登録日: 2017-08-29, 公開日: 2018-12-12, 最終更新日: 2024-05-15)

主引用文献

Viegas, A.,Dollinger, P.,Verma, N.,Kubiak, J.,Viennet, T.,Seidel, C.A.M.,Gohlke, H.,Etzkorn, M.,Kovacic, F.,Jaeger, K.E.
Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation.
Sci Rep, 10:3578-3578, 2020

PubMed Abstract: Folding and cellular localization of many proteins of Gram-negative bacteria rely on a network of chaperones and secretion systems. Among them is the lipase-specific foldase Lif, a membrane-bound steric chaperone that tightly binds (K = 29 nM) and mediates folding of the lipase LipA, a virulence factor of the pathogenic bacterium P. aeruginosa. Lif consists of five-domains, including a mini domain MD1 essential for LipA folding. However, the molecular mechanism of Lif-assisted LipA folding remains elusive. Here, we show in in vitro experiments using a soluble form of Lif (sLif) that isolated MD1 inhibits sLif-assisted LipA activation. Furthermore, the ability to activate LipA is lost in the variant sLif, in which the evolutionary conserved amino acid Y99 from helix α1 of MD1 is mutated to alanine. This coincides with an approximately three-fold reduced affinity of the variant to LipA together with increased flexibility of sLif in the complex as determined by polarization-resolved fluorescence spectroscopy. We have solved the NMR solution structures of P. aeruginosa MD1 and variant MD1 revealing a similar fold indicating that a structural modification is likely not the reason for the impaired activity of variant sLif. Molecular dynamics simulations of the sLif:LipA complex in connection with rigidity analyses suggest a long-range network of interactions spanning from Y99 of sLif to the active site of LipA, which might be essential for LipA activation. These findings provide important details about the putative mechanism for LipA activation and point to a general mechanism of protein folding by multi-domain steric chaperones.

PubMed: 32107397
DOI: 10.1038/s41598-020-60093-4

実験手法

SOLUTION NMR