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Cry-EM and Single Particle Reconstruction



DESCRIPTION OF LAB RESEARCH WORK
We are interested in determining 3D structures of macromolecular protein complexes to understand their function and mechanism. We employ Cryo-Electron Microscopy with single particle reconstruction as our main tool for structural determination. Our laboratory is equipped with advanced Electron Microscopes and high-speed computing clusters for routine single particle reconstruction work. We are mainly interested in the following research areas:
  1. Protein translocation across cellular membranes
  2. Organellar ribosomes
  3. Protein secretion
  4. ​Drug discovery and multidrug resistance in Mycobacteria


LAB MEMBERS
LEAD PI
Shashi Bhushan
Associate Professor

Email: sbhushan@ntu.edu.sg
Phone: (65) 6592 3673
Office: SBS-02N-01
Ambuj Kumar Kushwaha
Research Fellow

Email: kushwaha@ntu.edu.sg​
Jian Sun
Research Fellow

Email: jsun@ntu.edu.sg​


CURRENT PROJECTS
  1. Structural investigation of organellar ribosomes
  2. Structural and biochemical investigation of potential targets in Mycobacteria

PUBLICATIONS
  • Kumar V, Chen Y, Ero R, Ahmed T, Tan J, Li Z, et al. Structure of BipA in GTP form bound to the ratcheted ribosome. Proceedings of the National Academy of Sciences of the United States of America. 2015;112(35):10944-9.
  • Tsai YCC, Lapina MC, Bhushan S, Mueller-Cajar O. Identification and characterization of multiple rubisco activases in chemoautotrophic bacteria. Nature Communications. 2015;6.
  • Ahmed T, Yin Z, Bhushan S. Cryo-EM structure of the large subunit of the spinach chloroplast ribosome. Scientific Reports. 2016;6.
  • Singh D, Sielaff H, Sundararaman L, Bhushan S, Grüber G. The stimulating role of subunit F in ATPase activity inside the A1-complex of the Methanosarcina mazei Gö1 A1AO ATP synthase. Biochimica et Biophysica Acta - Bioenergetics. 2016;1857(2):177-87.
  • Ahmed T, Shi J, Bhushan S. Unique localization of the plastid-specific ribosomal proteins in the chloroplast ribosome small subunit provides mechanistic insights into the chloroplastic translation. Nucleic Acids Research. 2017;45(14):8581-95.
  • Belousoff MJ, Eyal Z, Radjainia M, Ahmed T, Bamert RS, Matzov D, et al. Structural basis for linezolid binding site rearrangement in the Staphylococcus aureus ribosome. mBio. 2017;8(3).
  • Gong Q, Long Z, Zhong FL, Teo DET, Jin Y, Yin Z, et al. Structural basis of RIP2 activation and signaling. Nature Communications. 2018;9(1).
  • Mishra S, Ahmed T, Tyagi A, Shi J, Bhushan S. Structures of Mycobacterium smegmatis 70S ribosomes in complex with HPF, tmRNA, and P-tRNA. Scientific Reports. 2018;8(1).
  • Pan A, Balakrishna AM, Nartey W, Kohlmeier A, Dip PV, Bhushan S, et al. Atomic structure and enzymatic insights into the vancomycin-resistant Enterococcus faecalis (V583) alkylhydroperoxide reductase subunit C. Free Radical Biology and Medicine. 2018;115:252-65.
  • Tharakaraman K, Watanabe S, Chan KR, Huan J, Subramanian V, Chionh YH, et al. Rational Engineering and Characterization of an mAb that Neutralizes Zika Virus by Targeting a Mutationally Constrained Quaternary Epitope. Cell Host and Microbe. 2018;23(5):618-27.e6.
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