The crystal structure of two macrolide glycosyltransferases provides a blueprint for host cell antibiotic immunity

Glycosylation of macrolide antibiotics confers host cell immunity from endogenous and exogenous agents. The Streptomyces antibioticus glycosyltransferases, OleI and OleD, glycosylate and inactivate oleandomycin and diverse macrolides including erythromycin, respectively. The structure of these enzyme–ligand complexes, in tandem with kinetic analysis of site-directed variants, provide insight into the interaction of macrolides with their synthetic apparatus. Erythromycin binds to OleD and the 23S RNA of its target ribosome in the same conformation and, although the antibiotic contains a large number of polar groups, its interaction with these macromolecules is primarily through hydrophobic contacts. Erythromycin and oleandomycin, when bound to OleD and OleI, respectively, adopt different conformations, reflecting a subtle effect on sugar positioning by virtue of a single change in the macrolide backbone. The data reported here provide structural insight into the mechanism of resistance to both endogenous and exogenous antibiotics, and will provide a platform for the future redesign of these catalysts for antibiotic remodelling.

[1]  C. Ford,et al.  Structure of a flavonoid glucosyltransferase reveals the basis for plant natural product modification , 2006 .

[2]  Bernard Henrissat,et al.  An evolving hierarchical family classification for glycosyltransferases. , 2003, Journal of molecular biology.

[3]  J. Errey,et al.  Probing the breadth of macrolide glycosyltransferases: in vitro remodeling of a polyketide antibiotic creates active bacterial uptake and enhances potency. , 2005, Journal of the American Chemical Society.

[4]  J. Salas,et al.  Biosynthesis of the Macrolide Oleandomycin by Streptomyces antibioticus , 1995, The Journal of Biological Chemistry.

[5]  R. Dixon,et al.  Crystal Structures of a Multifunctional Triterpene/Flavonoid Glycosyltransferase from Medicago truncatula , 2005, The Plant Cell Online.

[6]  C. Méndez,et al.  Role of glycosylation and deglycosylation in biosynthesis of and resistance to oleandomycin in the producer organism, Streptomyces antibioticus , 1992, Journal of bacteriology.

[7]  Pedro M Alzari,et al.  Crystal structure of glycogen synthase: homologous enzymes catalyze glycogen synthesis and degradation , 2004, The EMBO journal.

[8]  Kevin Cowtan,et al.  research papers Acta Crystallographica Section D Biological , 2005 .

[9]  C. Sander,et al.  Protein structure comparison by alignment of distance matrices. , 1993, Journal of molecular biology.

[10]  Wei Lu,et al.  Structure of the TDP-epi-vancosaminyltransferase GtfA from the chloroeremomycin biosynthetic pathway , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[11]  R M Esnouf,et al.  An extensively modified version of MolScript that includes greatly enhanced coloring capabilities. , 1997, Journal of molecular graphics & modelling.

[12]  J. Salas,et al.  Glycosylation of Macrolide Antibiotics , 2000, The Journal of Biological Chemistry.

[13]  S. Walker,et al.  Crystal structure of the MurG:UDP-GlcNAc complex reveals common structural principles of a superfamily of glycosyltransferases , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Yigong Shi,et al.  The 1.9 Å crystal structure of Escherichia coli MurG, a membrane‐associated glycosyltransferase involved in peptidoglycan biosynthesis , 2000, Protein science : a publication of the Protein Society.

[15]  Gregor Blaha,et al.  Structures of MLSBK Antibiotics Bound to Mutated Large Ribosomal Subunits Provide a Structural Explanation for Resistance , 2005, Cell.

[16]  D. Denning,et al.  In vitro activity of a new echinocandin, LY303366, and comparison with fluconazole, flucytosine and amphotericin B against Candida species. , 2001, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[17]  M. Palcic,et al.  A continuous spectrophotometric assay for glycosyltransferases. , 1994, Analytical biochemistry.

[18]  S. Walker,et al.  Remarkable structural similarities between diverse glycosyltransferases. , 2002, Chemistry & biology.

[19]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[20]  C. Walsh,et al.  Crystal structure of vancosaminyltransferase GtfD from the vancomycin biosynthetic pathway: interactions with acceptor and nucleotide ligands. , 2004, Biochemistry.

[21]  C. Méndez,et al.  Altering the glycosylation pattern of bioactive compounds. , 2001, Trends in biotechnology.

[22]  C. Méndez,et al.  Two glycosyltransferases and a glycosidase are involved in oleandomycin modification during its biosynthesis by Streptomyces antibioticus , 1998, Molecular microbiology.

[23]  Alexei G. Sankovski,et al.  Special report on emissions scenarios : a special report of Working group III of the Intergovernmental Panel on Climate Change , 2000 .

[24]  Randy J Read,et al.  Electronic Reprint Biological Crystallography Likelihood-enhanced Fast Translation Functions Biological Crystallography Likelihood-enhanced Fast Translation Functions , 2022 .

[25]  A. Kubo,et al.  Alteration of sugar donor specificities of plant glycosyltransferases by a single point mutation. , 2004, Archives of biochemistry and biophysics.

[26]  C. Méndez,et al.  Characterization of a Streptomyces antibioticus gene cluster encoding a glycosyltransferase involved in oleandomycin inactivation. , 1993, Gene.

[27]  Anastassis Perrakis,et al.  Automated protein model building combined with iterative structure refinement , 1999, Nature Structural Biology.

[28]  N. H. Ravindranath,et al.  Land Use, Land-Use Change, and Forestry: A Special Report of the Intergovernmental Panel on Climate Change , 2000 .

[29]  George M Sheldrick,et al.  Substructure solution with SHELXD. , 2002, Acta crystallographica. Section D, Biological crystallography.

[30]  R. Campbell,et al.  The structure of UDP-N-acetylglucosamine 2-epimerase reveals homology to phosphoglycosyl transferases. , 2000, Biochemistry.

[31]  S. Withers,et al.  Engineering of glycosidases and glycosyltransferases. , 2006, Current opinion in chemical biology.

[32]  G. Murshudov,et al.  Refinement of macromolecular structures by the maximum-likelihood method. , 1997, Acta crystallographica. Section D, Biological crystallography.

[33]  C. Walsh,et al.  Structure of the UDP-glucosyltransferase GtfB that modifies the heptapeptide aglycone in the biosynthesis of vancomycin group antibiotics. , 2001, Structure.

[34]  Benjamin G Davis,et al.  Structural dissection and high-throughput screening of mannosylglycerate synthase , 2005, Nature Structural &Molecular Biology.

[35]  S. Withers,et al.  The Donor Subsite of Trehalose-6-phosphate Synthase , 2004, Journal of Biological Chemistry.