Cry-EM and Single Particle Reconstruction


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


Shashi BhushanLEAD PI
Shashi Bhushan
Associate Professor

Email: [email protected]
Phone: (65) 6592 3673
Office: SBS-02N-01
Ambuj Kumar Kushwaha
Research Fellow

Email: [email protected]
Jian Sun
Research Fellow

Email: [email protected]

1. Structure, function, and pharmacology of membrane proteins

  • Human voltage-gated potassium ion channels
  • Human calcium-gated potassium ion channels

2. Infectious diseases

  • Structural pharmacology of Malaria parasite
  • Structure of therapeutical anti-viral antibodies

3. Structure, function, and regulation of protein translation machinery

4. Structure and function of cellular machineries

Full list of publication can be found here.

  • Ong, S. T., Tyagi, A., Chandy, K. G., and Bhushan, S. (2022) Mechanisms Underlying C-type Inactivation in Kv Channels: Lessons From Structures of Human Kv1.3 and Fly Shaker-IR Channels. Front Pharmacol 13, 924289 (
  • Tyagi, A., Ahmed, T., Jian, S., Bajaj, S., Ong, S. T., Goay, S. S. M., Zhao, Y., Vorobyov, I., Tian, C., Chandy, K. G., and Bhushan, S. (2022) Rearrangement of a unique Kv1.3 selectivity filter conformation upon binding of a drug. Proc Natl Acad Sci USA 119 e2113536119.(
  • Tyagi, A., Ahmed, T., Shi, J., and Bhushan, S. (2020) A complex between the Zika virion and the Fab of a broadly cross-reactive neutralizing monoclonal antibody revealed by cryo-EM and single particle analysis at 4.1 A resolution. J Struct Biol X 4, 100028 (
  • Kushwaha, A. K., and Bhushan, S. (2020) Unique structural features of the Mycobacterium ribosome. Prog Biophys Mol Biol 152, 15-24 (
  • Gopal, P., Sarathy, J. P., Yee, M., Ragunathan, P., Shin, J., Bhushan, S., Zhu, J., Akopian, T., Kandror, O., Lim, T. K., Gengenbacher, M., Lin, Q., Rubin, E. J., Gruber, G., and Dick, T. (2020) Pyrazinamide triggers degradation of its target aspartate decarboxylase. Nat Commun 11, 1661(
  • Tsai, Y. C., Ye, F., Liew, L., Liu, D., Bhushan, S., Gao, Y. G., and Mueller-Cajar, O. (2020) Insights into the mechanism and regulation of the CbbQO-type Rubisco activase, a MoxR AAA+ ATPase. Proc Natl Acad Sci USA 117, 381-387 (
  • Tharakaraman, K., Watanabe, S., Chan, K. R., Huan, J., Subramanian, V., Chionh, Y. H., Raguram, A., Quinlan, D., McBee, M., Ong, E. Z., Gan, E. S., Tan, H. C., Tyagi, A., Bhushan, S., Lescar, J., Vasudevan, S. G., Ooi, E. E., and Sasisekharan, R. (2018) Rational Engineering and Characterization of an mAb that Neutralizes Zika Virus by Targeting a Mutationally Constrained Quaternary Epitope. Cell Host Microbe 23, 618-627.e616 (
  • Pan, A., Balakrishna, A. M., Nartey, W., Kohlmeier, A., Dip, P. V., Bhushan, S., and Grüber, G. (2018) Atomic structure and enzymatic insights into the vancomycin-resistant Enterococcus faecalis (V583) alkylhydroperoxide reductase subunit C. Free Radic Biol Med 115, 252-265 (
  • Mishra, S., Ahmed, T., Tyagi, A., Shi, J., and Bhushan, S. (2018) Structures of Mycobacterium smegmatis 70S ribosomes in complex with HPF, tmRNA, and P-tRNA. Sci Rep 8, 13587 (
  • Gong, Q., Long, Z., Zhong, F. L., Teo, D. E. T., Jin, Y., Yin, Z., Boo, Z. Z., Zhang, Y., Zhang, J., Yang, R., Bhushan, S., Reversade, B., Li, Z., and Wu, B. (2018) Structural basis of RIP2 activation and signaling. Nat Commun 9 (