Structural remodelling of the carbon–phosphorus lyase machinery by a dual ABC ATPase

DOI: 10.1038/s41467-023-36604-y

Authors: Søren K. Amstrup (Aarhus University) , Sui Ching Ong (Aarhus University) , Nicholas Sofos (Aarhus University) , Jesper L. Karlsen (Aarhus University) , Ragnhild B. Skjerning (Aarhus University) , Thomas Boesen (Aarhus University) , Jan J. Enghild (Aarhus University) , Bjarne Hove-Jensen (Aarhus University) , Ditlev E. Brodersen (Aarhus University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 14

State: Published (Approved)
Published: February 2023
Diamond Proposal Number(s): 21404

Abstract: In Escherichia coli, the 14-cistron phn operon encoding carbon-phosphorus lyase allows for utilisation of phosphorus from a wide range of stable phosphonate compounds containing a C-P bond. As part of a complex, multi-step pathway, the PhnJ subunit was shown to cleave the C-P bond via a radical mechanism, however, the details of the reaction could not immediately be reconciled with the crystal structure of a 220 kDa PhnGHIJ C-P lyase core complex, leaving a significant gap in our understanding of phosphonate breakdown in bacteria. Here, we show using single-particle cryogenic electron microscopy that PhnJ mediates binding of a double dimer of the ATP-binding cassette proteins, PhnK and PhnL, to the core complex. ATP hydrolysis induces drastic structural remodelling leading to opening of the core complex and reconfiguration of a metal-binding and putative active site located at the interface between the PhnI and PhnJ subunits.

Journal Keywords: Bacterial structural biology; Cryoelectron microscopy; Enzyme mechanisms; Multienzyme complexes; Soil microbiology

Diamond Keywords: Enzymes; Bacteria

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios IV-Titan Krios IV at Diamond

Other Facilities: EMBION

Added On: 01/03/2023 10:36

Documents:
s41467-023-36604-y.pdf

Discipline Tags:

Structural biology Life Sciences & Biotech

Technical Tags:

Microscopy Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)