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Structural Basis of Pullulanase Membrane Binding and Secretion Revealed by X-Ray Crystallography, Molecular Dynamics and Biochemical Analysis.
- Source :
-
Structure (London, England : 1993) [Structure] 2016 Jan 05; Vol. 24 (1), pp. 92-104. Date of Electronic Publication: 2015 Dec 10. - Publication Year :
- 2016
-
Abstract
- The Klebsiella lipoprotein pullulanase (PulA) is exported to the periplasm, triacylated, and anchored via lipids in the inner membrane (IM) prior to its transport to the bacterial surface through a type II secretion system (T2SS). X-Ray crystallography and atomistic molecular dynamics (MD) simulations of PulA in a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) model membrane provided an unprecedented molecular view of an N-terminal unstructured tether and the IM lipoprotein retention signal, and revealed novel interactions with the IM via N-terminal immunoglobulin-like domains in PulA. An efficiently secreted nonacylated variant (PulANA) showed similar peripheral membrane association during MD simulations, consistent with the binding of purified PulANA to liposomes. Remarkably, combined X-ray, MD, and functional studies identified a novel subdomain, Ins, inserted in the α-amylase domain, which is required for PulA secretion. Available data support a model in which PulA binding to the IM promotes interactions with the T2SS, possibly via the Ins subdomain.<br /> (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Subjects :
- Amino Acid Sequence
Bacterial Proteins metabolism
Binding Sites
Crystallography, X-Ray
Exocytosis
Glycoside Hydrolases metabolism
Klebsiella enzymology
Lipid Bilayers chemistry
Lipid Bilayers metabolism
Lipoproteins metabolism
Membrane Proteins metabolism
Molecular Sequence Data
Protein Binding
Bacterial Proteins chemistry
Cell Membrane metabolism
Glycoside Hydrolases chemistry
Molecular Dynamics Simulation
Subjects
Details
- Language :
- English
- ISSN :
- 1878-4186
- Volume :
- 24
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Structure (London, England : 1993)
- Publication Type :
- Academic Journal
- Accession number :
- 26688215
- Full Text :
- https://doi.org/10.1016/j.str.2015.10.023