2YOC

Crystal structure of PulA from Klebsiella oxytoca

Summary for 2YOC

• Entry DOI: 10.2210/pdb2yoc/pdb
• Descriptor: PULLULANASE, CALCIUM ION, SULFATE ION, ... (4 entities in total)
• Functional Keywords: hydrolase, glycosyl hydrolase, type 2 secretion systems
• Biological source: KLEBSIELLA OXYTOCA
• Cellular location: Cell membrane ; Lipid-anchor : P07206
• Total number of polymer chains: 2
• Total formula weight: 238215.33

Authors

Francetic, O.,Mechaly, A.E.,Tello-Manigne, D.,Buschiazzo, A.,Bernarde, C.,Nadeau, N.,Pugsley, A.P.,Alzari, P.M. (deposition date: 2012-10-23, release date: 2013-11-06, Last modification date: 2024-11-06)

Primary citation

East, A.,Mechaly, A.E.,Huysmans, G.H.M.,Bernarde, C.,Tello-Manigne, D.,Nadeau, N.,Pugsley, A.P.,Buschiazzo, A.,Alzari, P.M.,Bond, P.J.,Francetic, O.
Structural Basis of Pullulanase Membrane Binding and Secretion Revealed by X-Ray Crystallography, Molecular Dynamics and Biochemical Analysis
Structure, 24:92-, 2016

PubMed 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.

PubMed: 26688215
DOI: 10.1016/J.STR.2015.10.023

Experimental method

X-RAY DIFFRACTION (2.88 Å)