Ali Kermani Email & Phone Number

Rockville, Maryland, Us

Memphis, Tennessee, Us

• Expressing mammalian proteins using cell culture (HEK 293 cells).• Optimizing the expression level and stability of hard-to-express proteins for structural studies using cryo-electron microscopy (cryo-EM).• Protein purification using affinity, ion exchange (IEX) and size exclusion chromatography (SEC) using AKTA systems.• Solving the high-resolution structures of membrane proteins, protein complexes, and protein-small molecules using cryo-EM.• Cryo-EM sample preparation, grid clipping and freezing, screening and data collection by Talos Arctica and Krios using Serial EM and EPU software. • Cryo-EM data processing using CryoSPARC and Relion and model building using Coot and Phenix. Visualizing the 3D structures using Pymol and ChimeraX. • Characterizing purified proteins using analytical chromatography techniques such as SEC, FSEC and DLS. • Supervising postdoctoral researchers.

Ann Arbor, Michigan, Us

• Involved in setting up the structural biology part of the lab from beginning. • Overexpression, purification and isolation of a wide range of soluble and transmembrane proteins, such as Small Multidrug Resistance (SMR) family of transmembrane proteins, K+ channels and Mg2+ transporters, using both prokaryotic and yeast expression systems. • Purification is performed using affinity chromatography, ion exchange chromatography and size exclusion chromatography run by AKTA and other automated systems. • Troubleshooting protein production in the case of challenging proteins. • Biochemical and biophysical characterization of purified proteins using isothermal titration calorimetry (ITC), FPLC, size exclusion chromatography (SEC) and small angle x-ray scattering (SAXS). • Structural determination of SMR proteins using X-ray crystallography, from crystal generation, data collection to model building. • Developing novel crystallography chaperones, such as Monobodies and Nanobodies for structural studies of target membrane proteins. • Studying the role of K+ channels on biofilm formation. • Supervising postgraduate and undergraduate students involved in these projects.

Birmingham, Al, Us

Biophysical and structural studies of outer-membrane proteins in M. tuberculosis X-ray structural determination of Hyaluronoglucosaminidase from Streptococcus pyogenes phage H4489A

Birmingham, West Midlands, Gb

My PhD thesis was focused on biophysical and biochemical characterization of proteins involved in capsule biosynthesis in Mycobacterium tuberculosis in the research group of Dr. Klaus Fütterer and Professor Gurdyal Besra at the Institute of Microbiology and Infection, University of Birmingham, United Kingdom. The cell wall in Mycobacterium tuberculosis, the causative agent of TB, is made of several layers which protect the bacterium against antibiotics and are involved in infection. Studying its biosynthetic pathway would enable us to develop new antibiotics to control and eliminate the TB infection.My PhD project involved:- Cloning of mycobacterial proteins TreS, Pep2 (MaK), GlgE, Rv3032, Rv3031, GlgA and GlgC, involved in α-glucan biosynthesis in mycobacteria- Protein overexpression and purification from E. coli and M. smegmatis cell extracts. - Developing and performing enzymatic assays to measure activity of (a subset) of these proteins- Protein crystallization in sparse matrix and systematic screens by sitting drop vapour diffusion. Co-crystallisation with ligands.- Probing solution behaviour of proteins by analytical ultracentrifugation in sedimentation velocity and equilibrium mode.- Probing protein-protein interactions by isothermal titration calorimetry and size exclusion chromatography.- X-ray diffraction data collection at synchrotron beamlines.- Structure solution and refinement based on X-ray diffraction data.

Newcastle Upon Tyne, Tyne And Wear, Gb

- Purifying LpoA, a membrane protein involved in peptidoglycan biosynthesis regulation, in order to study its putative interactions with other proteins- Measuring the rate of peptidoglycan incorporation into the sacculus in vivo by performing incorporation assay- Analysis of peptidoglycan structure of different strains using HPLC- Studying the sacculi of different strains using electron-microscope (TEM) and other imaging techniques

Newcastle Upon Tyne, Gb

Discovery and purification of antibiotics from actinomycetes. This involved:- Growth of filamentous actinomycetes, notably streptomycetes isolated from hyper-saline Atacama Desert soil on media designed to promote the production of bioactive compounds. - Determining evolutionary position of the representative strains in the 16S rRNA actinobacterial gene tree.- Screening strains from distinct phyletic lines of the 16S rRNA actinobacterial gene tree using a panel of clinically significant microorganisms in agar plug assays. - Determining the mode of action of detected bioactive compounds in agar plug assays using Bacillus subtilis reporter strains.- Preliminary analysis of the structure of selected novel bioactive agents using high-performance liquid chromatography and mass spectrometry.

Phd, Structural Biology

Masters, Industrial And Commercial Biotechnology