Structural insights into the bacterial carbon-phosphorus lyase machinery.
Phosphorus is required for all life and microorganisms can extract it from their environment through several metabolic pathways. When phosphate is in limited supply, some bacteria are able to use phosphonate compounds, which require specialized enzymatic machinery to break the stable carbon-phosphorus (C-P) bond. Despite its importance, the details of how this machinery catabolizes phosphonates remain unknown. Here we determine the crystal structure of the 240-kilodalton Escherichia coli C-P lyase core complex (PhnG-PhnH-PhnI-PhnJ; PhnGHIJ), and show that it is a two-fold symmetric hetero-octamer comprising an intertwined network of subunits with unexpected self-homologies. It contains two potential active sites that probably couple phosphonate compounds to ATP and subsequently hydrolyse the C-P bond. We map the binding site of PhnK on the complex using electron microscopy, and show that it binds to a conserved insertion domain of PhnJ. Our results provide a structural basis for understanding microbial phosphonate breakdown.
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark.
Paulina
P
Seweryn
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark.
Lan Bich
LB
Van
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark.
Morten
M
Kjeldgaard
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Christopher J
CJ
Russo
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Lori A
LA
Passmore
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark.
Bjarne
B
Hove-Jensen
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark.
Bjarne
B
Jochimsen
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark.
Ditlev E
DE
Brodersen
D000255:Adenosine Triphosphate / Q000378:metabolism; D001665:Binding Sites; D055162:Biocatalysis; D002244:Carbon / Q000737:chemistry / Q000378:metabolism; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D006868:Hydrolysis; D007501:Iron / Q000737:chemistry / Q000378:metabolism; D008190:Lyases / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008958:Models, Molecular; D063065:Organophosphonates / Q000378:metabolism; D010758:Phosphorus / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D013455:Sulfur / Q000737:chemistry / Q000378:metabolism
D029968:Escherichia coli Proteins; D063065:Organophosphonates; C000601555:PhnK protein, E coli; D021122:Protein Subunits; D010758:Phosphorus; D013455:Sulfur; D002244:Carbon; D000255:Adenosine Triphosphate; D007501:Iron; D008190:Lyases; C051328:carbon-phosphorus lyase