PDBsum entry 2yoc

Hydrolase

PDB id: 2yoc

Contents

Protein chains

1051 a.a.

Ligands

SO4 ×14

Metals

_CA ×8

Waters ×308

PDB id:

2yoc

Name:

Hydrolase

Title:

Crystal structure of pula from klebsiella oxytoca

Structure:

Pullulanase. Chain: a, b. Fragment: residues 21-1089. Synonym: alpha-dextrin endo-1,6-alpha-glucosidase, pullulan 6- glucanohydrolase. Engineered: yes

Source:

Klebsiella oxytoca. Organism_taxid: 571. Strain: unf5023. Expressed in: escherichia coli b. Expression_system_taxid: 37762.

Resolution:

2.88Å R-factor: 0.173 R-free: 0.213

Authors:

O.Francetic,A.E.Mechaly,D.Tello-Manigne,A.Buschiazzo,C.Bernarde, N.Nadeau,A.P.Pugsley,P.M.Alzari

Key ref:

A.East et al. (2016). Structural Basis of Pullulanase Membrane Binding and Secretion Revealed by X-Ray Crystallography, Molecular Dynamics and Biochemical Analysis. Structure, 24, 92. PubMed id: 26688215 DOI: 10.1016/j.str.2015.10.023

Date:

23-Oct-12 Release date: 06-Nov-13

Protein chains

P07206  (PULA_KLEPN) -  Pullulanase from Klebsiella pneumoniae

Seq: 1090 a.a.

Struc: 1051 a.a.

Enzyme reactions

• Enzyme class: E.C.3.2.1.41 - pullulanase.
• Reaction: Hydrolysis of (1->6)-alpha-D-glucosidic linkages in pullulan and in amylopectin and glycogen, and the alpha- and beta-limit dextrins of amylopectin and glycogen.

DOI no: 10.1016/j.str.2015.10.023

Structure 24:92 (2016)

PubMed id: 26688215

Structural Basis of Pullulanase Membrane Binding and Secretion Revealed by X-Ray Crystallography, Molecular Dynamics and Biochemical Analysis.

A.East, A.E.Mechaly, G.H.Huysmans, C.Bernarde, D.Tello-Manigne, N.Nadeau, A.P.Pugsley, A.Buschiazzo, P.M.Alzari, P.J.Bond, O.Francetic.

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.