The structure of a calcium-dependent phosphoinositide-specific phospholipase C from Pseudomonas sp. 62186, the first from a Gram-negative bacterium

TY - JOUR

T1 - The structure of a calcium-dependent phosphoinositide-specific phospholipase C from Pseudomonas sp. 62186, the first from a Gram-negative bacterium

AU - Moroz, Olga V.

AU - Blagova, Elena

AU - Lebedev, Andrey A.

AU - Nørgaard, Allan

AU - Segura, Dorotea R.

AU - Blicher, Thomas H.

AU - Brask, Jesper

AU - Wilson, Keith S.

N1 - ©2017 International Union of Crystallography. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details.

PY - 2017/1/1

Y1 - 2017/1/1

N2 - Bacterial phosphoinositide-specific phospholipases C (PI-PLCs) are the smallest members of the PI-PLC family, which includes much larger mammalian enzymes responsible for signal transduction as well as enzymes from protozoan parasites, yeast and plants. Eukaryotic PI-PLCs have calcium in the active site, but this is absent in the known structures of Gram-positive bacteria, where its role is instead played by arginine. In addition to their use in a number of industrial applications, the bacterial enzymes attract special interest because they can serve as convenient models of the catalytic domains of eukaryotic enzymes for in vitro activity studies. Here, the structure of a PI-PLC from Pseudomonas sp. 62186 is reported, the first from a Gram-negative bacterium and the first of a native bacterial PI-PLC with calcium present in the active site. Solution of the structure posed particular problems owing to the low sequence identity of available homologous structures. Its dependence on calcium for catalysis makes this enzyme a better model for studies of the mammalian PI-PLCs than the previously used calcium-independent bacterial PI-PLCs.The structure of the calcium-dependent phosphoinositide-specific phospholipase C from Pseudomonas sp. 62186 is described, the first from a Gram-negative bacterium. Structure solution from a low-sequence-identity homology model is described in detail.

AB - Bacterial phosphoinositide-specific phospholipases C (PI-PLCs) are the smallest members of the PI-PLC family, which includes much larger mammalian enzymes responsible for signal transduction as well as enzymes from protozoan parasites, yeast and plants. Eukaryotic PI-PLCs have calcium in the active site, but this is absent in the known structures of Gram-positive bacteria, where its role is instead played by arginine. In addition to their use in a number of industrial applications, the bacterial enzymes attract special interest because they can serve as convenient models of the catalytic domains of eukaryotic enzymes for in vitro activity studies. Here, the structure of a PI-PLC from Pseudomonas sp. 62186 is reported, the first from a Gram-negative bacterium and the first of a native bacterial PI-PLC with calcium present in the active site. Solution of the structure posed particular problems owing to the low sequence identity of available homologous structures. Its dependence on calcium for catalysis makes this enzyme a better model for studies of the mammalian PI-PLCs than the previously used calcium-independent bacterial PI-PLCs.The structure of the calcium-dependent phosphoinositide-specific phospholipase C from Pseudomonas sp. 62186 is described, the first from a Gram-negative bacterium. Structure solution from a low-sequence-identity homology model is described in detail.

KW - calcium-dependent

KW - Gram-negative bacterium

KW - phosphoinositol-specific

KW - phospholipase C

KW - Pseudomonas sp. 62186

KW - X-ray structure

UR - http://www.scopus.com/inward/record.url?scp=85008237706&partnerID=8YFLogxK

U2 - 10.1107/S2059798316019616

DO - 10.1107/S2059798316019616

M3 - Article

AN - SCOPUS:85008237706

SN - 2059-7983

VL - 73

SP - 32

EP - 44

JO - Acta crystallographica. Section D, Structural biology

JF - Acta crystallographica. Section D, Structural biology

IS - 1

ER -