Cryo-EM structure of arabinosyltransferase EmbB from Mycobacterium smegmatis

Yong Zi Tan; José Rodrigues; James E Keener; Ruixiang Blake Zheng; Richard Brunton; Brian Kloss; Sabrina I Giacometti; Ana L Rosário; Lei Zhang; Michael Niederweis; Oliver B Clarke; Todd L Lowary; Michael T Marty; Margarida Archer; Clinton S Potter; Bridget Carragher; Filippo Mancia

doi:10.1038/s41467-020-17202-8

Cryo-EM structure of arabinosyltransferase EmbB from Mycobacterium smegmatis

Nat Commun. 2020 Jul 7;11(1):3396. doi: 10.1038/s41467-020-17202-8.

Authors

Yong Zi Tan^{1

2}, José Rodrigues³, James E Keener⁴, Ruixiang Blake Zheng⁵, Richard Brunton⁵, Brian Kloss⁶, Sabrina I Giacometti¹, Ana L Rosário³, Lei Zhang⁷, Michael Niederweis⁷, Oliver B Clarke^{1

8}, Todd L Lowary^{5

9}, Michael T Marty^{4

10}, Margarida Archer³, Clinton S Potter^{2

11

12}, Bridget Carragher^{13

14

15}, Filippo Mancia¹⁶

Affiliations

¹ Department of Physiology and Cellular Biophysics, Columbia University, New York, NY, 10032, USA.
² National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, 10027, USA.
³ Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa (ITQB NOVA), 2780-157, Oeiras, Portugal.
⁴ Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ, 85721, USA.
⁵ Department of Chemistry, University of Alberta, Edmonton, Alberta, T6G 2G2, Canada.
⁶ Center on Membrane Protein Production and Analysis, New York Structural Biology Center, New York, NY, 10027, USA.
⁷ Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, 35294, USA.
⁸ Department of Anesthesiology, Columbia University, New York, NY, 10032, USA.
⁹ Institute of Biological Chemistry, Academia Sinica, Academia Road, Section 2, #128, Nangang, Taipei, 11529, Taiwan.
¹⁰ Bio5 Institute, University of Arizona, Tucson, AZ, 85721, USA.
¹¹ Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, 10027, USA.
¹² Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, 10032, USA.
¹³ National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, 10027, USA. bcarr@nysbc.org.
¹⁴ Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, 10027, USA. bcarr@nysbc.org.
¹⁵ Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, 10032, USA. bcarr@nysbc.org.
¹⁶ Department of Physiology and Cellular Biophysics, Columbia University, New York, NY, 10032, USA. fm123@cumc.columbia.edu.

Abstract

Arabinosyltransferase B (EmbB) belongs to a family of membrane-bound glycosyltransferases that build the lipidated polysaccharides of the mycobacterial cell envelope, and are targets of anti-tuberculosis drug ethambutol. We present the 3.3 Å resolution single-particle cryo-electron microscopy structure of Mycobacterium smegmatis EmbB, providing insights on substrate binding and reaction mechanism. Mutations that confer ethambutol resistance map mostly around the putative active site, suggesting this to be the location of drug binding.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Antitubercular Agents / pharmacology
Catalytic Domain
Cryoelectron Microscopy
Drug Resistance, Bacterial
Ethambutol / pharmacology
Lipids / chemistry
Mutation
Mycobacterium smegmatis / enzymology*
Mycobacterium tuberculosis / enzymology
Pentosyltransferases / chemistry*
Pentosyltransferases / ultrastructure*
Polysaccharides / chemistry
Protein Binding

Substances

Antitubercular Agents
Lipids
Polysaccharides
Ethambutol
EmbB protein, Mycobacterium tuberculosis
Pentosyltransferases

Abstract

Publication types

MeSH terms

Substances

Grants and funding