Research Overview:

We investigate the molecular mechanisms of bacterial protein synthesis and their links to antimicrobial resistance (AMR). We combine cryo-EM with biochemical and biophysical approaches to visualize macromolecular machines at near-atomic resolution.

A central focus is the structural and mechanistic characterization of bacterial ribosomes in complex with antibiotics, translation factors, and regulatory proteins — revealing how antibiotics inhibit translation, how bacteria evade them, and how resistance emerges. We also study cellular factors that modulate translation under stress conditions such as nutrient limitation and antibiotic exposure, connecting translational regulation to bacterial adaptation and persistence.

Our ultimate goal is to generate mechanistic insights that drive the development of next-generation antimicrobials and help address the global AMR crisis.

Cryo-EM Structure Determination Pipeline:

1. Sample Preparation – Purify the macromolecular complex and optimize buffer conditions for stability.

2. Vitrification – Flash-freeze sample in liquid ethane to preserve its native structure in a thin ice layer.

3. Data Collection – Acquire thousands of high-resolution micrographs using DED camera in a 200kV or 300kV cryo-microscope.

4. Particle Picking – Automatically or manually select particles of interest from micrographs.

5. 2D Classification – Group similar particle images to select good ones and remove junk/contaminants.

6. 3D Reconstruction – Generate an initial 3D density map using reference-based or ab-initio methods.

7. Refinement & Post-processing – Improve map resolution through iterative alignment and sharpening.

8. Model Building & Validation – Build, fit and refine atomic models into the density map using computational tools.

9. Deposition & Analysis – Submit structures to public databases (EMDB, PDB) and interpret biological insights.

Learn more about Cryo-EM