Journal Article

FZJ-2020-01347

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation

Viegas, A. ; Dollinger, P. ; Verma, N. ; Kubiak, J. ; Viennet, T. ; Seidel, C. A. M.Extern* ; Gohlke, H. (Corresponding author)FZJ* ; Etzkorn, M. (Corresponding author)FZJ* ; Kovacic, F.FZJ* ; Jaeger, K.-E. (Corresponding author)FZJ*

2020
Macmillan Publishers Limited, part of Springer Nature [London]

This record in other databases:      

Please use a persistent id in citations: http://hdl.handle.net/2128/24436  doi:10.1038/s41598-020-60093-4

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 (KD = 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 sLifY99A, 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 sLifY99A in the complex as determined by polarization-resolved fluorescence spectroscopy. We have solved the NMR solution structures of P. aeruginosa MD1 and variant MD1Y99A revealing a similar fold indicating that a structural modification is likely not the reason for the impaired activity of variant sLifY99A. 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.

---

Contributing Institute(s):

1. Jülich Supercomputing Center (JSC)
2. John von Neumann - Institut für Computing (NIC)
3. Strukturbiochemie (ICS-6)
4. Biotechnologie (IBG-1)
5. Institut für Molekulare Enzymtechnologie (HHUD) (IMET)

Research Program(s):

1. 511 - Computational Science and Mathematical Methods (POF3-511) (POF3-511)
2. Forschergruppe Gohlke (hkf7_20170501) (hkf7_20170501)
3. Conformational dynamics of the unbound lipase-specific foldase Lif (hdd16_20161101) (hdd16_20161101)
4. Analysis of the conformational changes during activation of lipase A by its foldase (hdd16_20171101) (hdd16_20171101)
5. 581 - Biotechnology (POF3-581) (POF3-581)

Appears in the scientific report 2020

---

Database coverage:
; ; ; ; BIOSIS Previews ; Clarivate Analytics Master Journal List ; Current Contents - Physical, Chemical and Earth Sciences ; DOAJ Seal ; Ebsco Academic Search ; IF < 5 ; JCR ; NCBI Molecular Biology Database ; PubMed Central ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Web of Science Core Collection ; Zoological Record

---