8S5B

Crystal structure of the sulfoquinovosyl binding protein (SmoF) from A. tumefaciens sulfo-SMO pathway in complex with SQOctyl ligand

This is a non-PDB format compatible entry.

Summary for 8S5B

• Entry DOI: 10.2210/pdb8s5b/pdb
• Descriptor: Sulfoquinovosyl glycerol-binding protein SmoF, [(2~{S},3~{S},4~{S},5~{R},6~{S})-6-octoxy-3,4,5-tris(oxidanyl)oxan-2-yl]methanesulfonic acid (3 entities in total)
• Functional Keywords: agrobacterium tumefaciens, sulfoquinovose, sulfosugar, solute binding protein, sulfoglycolysis, sulfo-smo pathway, sugar binding protein
• Biological source: Agrobacterium tumefaciens
• Total number of polymer chains: 1
• Total formula weight: 44431.27

Authors

Snow, A.J.D.,Sharma, M.,Davies, G.J. (deposition date: 2024-02-23, release date: 2024-04-17, Last modification date: 2024-05-08)

Primary citation

Arumapperuma, T.,Snow, A.J.D.,Lee, M.,Sharma, M.,Zhang, Y.,Lingford, J.P.,Goddard-Borger, E.D.,Davies, G.J.,Williams, S.J.
Capture-and-release of a sulfoquinovose-binding protein on sulfoquinovose-modified agarose.
Org.Biomol.Chem., 22:3237-3244, 2024

PubMed Abstract: The solute-binding protein (SBP) components of periplasmic binding protein-dependent ATP-binding cassette (ABC)-type transporters often possess exquisite selectivity for their cognate ligands. Maltose binding protein (MBP), the best studied of these SBPs, has been extensively used as a fusion partner to enable the affinity purification of recombinant proteins. However, other SBPs and SBP-ligand based affinity systems remain underexplored. The sulfoquinovose-binding protein SmoF, is a substrate-binding protein component of the ABC transporter cassette in involved in importing sulfoquinovose (SQ) and its derivatives for SQ catabolism. Here, we show that SmoF binds with high affinity to the octyl glycoside of SQ (octyl-SQ), demonstrating remarkable tolerance to extension of the anomeric substituent. The 3D X-ray structure of the SmoF·octyl-SQ complex reveals accommodation of the octyl chain, which projects to the protein surface, providing impetus for the synthesis of a linker-equipped SQ-amine using a thiol-ene reaction as a key step, and its conjugation to cyanogen bromide modified agarose. We demonstrate the successful capture and release of SmoF from SQ-agarose resin using SQ as competitive eluant, and selectivity for release other organosulfonates. We show that SmoF can be captured and purified from a cell lysate, demonstrating the utility of SQ-agarose in capturing SQ binding proteins from complex mixtures. The present work provides a pathway for development of 'capture-and-release' affinity resins for the discovery and study of SBPs.

PubMed: 38567495
DOI: 10.1039/d4ob00307a

Experimental method

X-RAY DIFFRACTION (1.8 Å)