5CVS

GlgE isoform 1 from Streptomyces coelicolor E423A mutant soaked in maltoheptaose

Summary for 5CVS

• Entry DOI: 10.2210/pdb5cvs/pdb
• Related PRD ID: PRD_900030
• Descriptor: Alpha-1,4-glucan:maltose-1-phosphate maltosyltransferase 1, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, ... (4 entities in total)
• Functional Keywords: hydrolase, alpha-glucan biosynthesis, glycoside hydrolase family 13_3, transferase
• Biological source: Streptomyces coelicolor
• Total number of polymer chains: 2
• Total formula weight: 154634.32

Authors

Rashid, A.M.,Syson, K.,Koliwer-Brandl, H.,van de Weerd, R.,Stevenson, C.E.M.,Batey, S.F.D.,Miah, F.,Alber, M.,Ioerger, T.R.,Chandra, G.,Appelmelk, B.J.,Nartowski, K.P.,Khimyak, Y.Z.,Lawson, D.M.,Jacobs, W.R.,Geurtsen, J.,Kalscheuer, R.,Bornemann, S. (deposition date: 2015-07-27, release date: 2016-08-17, Last modification date: 2024-01-10)

Primary citation

Syson, K.,Stevenson, C.E.,Miah, F.,Barclay, J.E.,Tang, M.,Gorelik, A.,Rashid, A.M.,Lawson, D.M.,Bornemann, S.
Ligand-bound structures and site-directed mutagenesis identify the acceptor and secondary binding sites of Streptomyces coelicolor maltosyltransferase GlgE.
J.Biol.Chem., 291:21531-21540, 2016

PubMed Abstract: GlgE is a maltosyltransferase involved in α-glucan biosynthesis in bacteria that has been genetically validated as a target for tuberculosis therapies. Crystals of the Mycobacterium tuberculosis enzyme diffract at low resolution so most structural studies have been with the very similar Streptomyces coelicolor GlgE isoform 1. Although the donor binding site for α-maltose 1-phosphate had been previously structurally defined, the acceptor site had not. Using mutagenesis, kinetics, and protein crystallography of the S. coelicolor enzyme, we have now identified the +1 to +6 subsites of the acceptor/product, which overlap with the known cyclodextrin binding site. The sugar residues in the acceptor subsites +1 to +5 are oriented such that they disfavor the binding of malto-oligosaccharides that bear branches at their 6-positions, consistent with the known acceptor chain specificity of GlgE. A secondary binding site remote from the catalytic center was identified that is distinct from one reported for the M. tuberculosis enzyme. This new site is capable of binding a branched α-glucan and is most likely involved in guiding acceptors toward the donor site because its disruption kinetically compromises the ability of GlgE to extend polymeric substrates. However, disruption of this site, which is conserved in the Streptomyces venezuelae GlgE enzyme, did not affect the growth of S. venezuelae or the structure of the polymeric product. The acceptor subsites +1 to +4 in the S. coelicolor enzyme are well conserved in the M. tuberculosis enzyme so their identification could help inform the design of inhibitors with therapeutic potential.

PubMed: 27531751
DOI: 10.1074/jbc.M116.748160

Experimental method

X-RAY DIFFRACTION (2.3 Å)