2LML

Solution NMR structure of holo acyl carrier protein from geobacter Metallireducens refined with nh rdcs, Northeast Structural Genomics consortium target gmr141

Replaces:  2KWMReplaces:  2KCIReplaces:  2KJS

Summary for 2LML

• Entry DOI: 10.2210/pdb2lml/pdb
• NMR Information: BMRB: 16860
• Descriptor: Putative acyl carrier protein, 4'-PHOSPHOPANTETHEINE (2 entities in total)
• Functional Keywords: structural genomics, psi-biology, protein structure initiative, northeast structural genomics consortium, nesg, transport protein
• Biological source: Geobacter metallireducens
• Total number of polymer chains: 1
• Total formula weight: 10382.80

Authors

Ramelot, T.A.,Smola, M.J.,Lee, H.,Zhao, L.,Ciccosanti, C.,Foote, E.L.,Hamilton, K.,Nair, R.,Rost, B.,Swapna, G.,Acton, T.B.,Xiao, R.,Everett, J.K.,Prestegard, J.H.,Montelione, G.T.,Kennedy, M.A.,Northeast Structural Genomics Consortium (NESG) (deposition date: 2011-12-05, release date: 2012-01-25, Last modification date: 2023-06-14)

Primary citation

Ramelot, T.A.,Smola, M.J.,Lee, H.W.,Ciccosanti, C.,Hamilton, K.,Acton, T.B.,Xiao, R.,Everett, J.K.,Prestegard, J.H.,Montelione, G.T.,Kennedy, M.A.
Solution structure of 4'-phosphopantetheine - GmACP3 from Geobacter metallireducens: a specialized acyl carrier protein with atypical structural features and a putative role in lipopolysaccharide biosynthesis.
Biochemistry, 50:1442-1453, 2011

PubMed Abstract: GmACP3 from Geobacter metallireducens is a specialized acyl carrier protein (ACP) whose gene, gmet_2339, is located near genes encoding many proteins involved in lipopolysaccharide (LPS) biosynthesis, indicating a likely function for GmACP3 in LPS production. By overexpression in Escherichia coli, about 50% holo-GmACP3 and 50% apo-GmACP3 were obtained. Apo-GmACP3 exhibited slow precipitation and non-monomeric behavior by (15)N NMR relaxation measurements. Addition of 4'-phosphopantetheine (4'-PP) via enzymatic conversion by E. coli holo-ACP synthase resulted in stable >95% holo-GmACP3 that was characterized as monomeric by (15)N relaxation measurements and had no indication of conformational exchange. We have determined a high-resolution solution structure of holo-GmACP3 by standard NMR methods, including refinement with two sets of NH residual dipolar couplings, allowing for a detailed structural analysis of the interactions between 4'-PP and GmACP3. Whereas the overall four helix bundle topology is similar to previously solved ACP structures, this structure has unique characteristics, including an ordered 4'-PP conformation that places the thiol at the entrance to a central hydrophobic cavity near a conserved hydrogen-bonded Trp-His pair. These residues are part of a conserved WDSLxH/N motif found in GmACP3 and its orthologs. The helix locations and the large hydrophobic cavity are more similar to medium- and long-chain acyl-ACPs than to other apo- and holo-ACP structures. Taken together, structural characterization along with bioinformatic analysis of nearby genes suggests that GmACP3 is involved in lipid A acylation, possibly by atypical long-chain hydroxy fatty acids, and potentially is involved in synthesis of secondary metabolites.

PubMed: 21235239
DOI: 10.1021/bi101932s

Experimental method

SOLUTION NMR