TY  - JOUR
AU  - Verma, Neha
AU  - Dollinger, Peter
AU  - Kovacic, Filip
AU  - Jaeger, Karl-Erich
AU  - Gohlke, Holger
TI  - The Membrane-Integrated Steric Chaperone Lif Facilitates Active Site Opening of Pseudomonas aeruginosa Lipase A
JO  - Journal of computational chemistry
VL  - 4
IS  - 6
SN  - 0192-8651
CY  - New York, NY [u.a.]
PB  - Wiley
M1  - FZJ-2019-05106
SP  - 500-512
PY  - 2020
AB  - Lipases are essential and widely used biocatalysts. Hence, the production of lipases requires a detailed understanding of the molecular mechanism of its folding and secretion. Lipase A from Pseudomonas aeruginosa, PaLipA, constitutes a prominent example that has additional relevance because of its role as a virulence factor in many diseases. PaLipA requires the assistance of a membrane‐integrated steric chaperone, the lipase‐specific foldase Lif, to achieve its enzymatically active state. However, the molecular mechanism of how Lif activates its cognate lipase has remained elusive. Here, we show by molecular dynamics simulations at the atomistic level and potential of mean force computations that Lif catalyzes the activation process of PaLipA by structurally stabilizing an intermediate PaLipA conformation, particularly a β‐sheet in the region of residues 17–30, such that the opening of PaLipA's lid domain is facilitated. This opening allows substrate access to PaLipA's catalytic site. A surprising and so far not fully understood aspect of our study is that the open state of PaLipA is unstable compared to the closed one according to our computational and in vitro biochemical results. We thus speculate that further interactions of PaLipA with the Xcp secretion machinery and/or components of the extracellular matrix contribute to the remaining activity of secreted PaLipA.
LB  - PUB:(DE-HGF)16
C6  - pmid:31618459
UR  - <Go to ISI:>//WOS:000490396800001
DO  - DOI:10.1002/jcc.26085
UR  - https://juser.fz-juelich.de/record/865808
ER  -