• Maintained protein crystallization facility infrastructure, screened crystallization conditions, and optimized hits for diffraction-quality crystal preparation; accessed multiple synchrotrons (NSLS, CLS, SSRL, ANSTO-MX, ESRF, ALS), collaborated with CROs for data collection, and performed structure solving and refinement.
• Prepared cryo-EM grids and coordinated shipping to NanoImaging Services for data collection; solved structures using Phenix, Chimera software using global and locally refined maps.
• Delivered 30+ X-ray and 28+ cryo-EM structures of oncology targets bound to natural ligands and inhibitors, elucidating key interactions and generating actionable design insights documented in comprehensive reports.
• Applied structure-based drug design (SBDD) and computational approaches (CADD) to guide strategic decision-making, accelerate hit-to-lead optimization, and achieve target potency and selectivity.
• Routinely participate in hit-finding initiatives with internal lead discovery teams (assay developments) and external CROs (AS-MS, covalent fragment libraries).
• Executed biophysical assays (nanoDSF) to assess protein thermal stability with ligands (Tm measurements), identifying stabilizers and aggregators to triage false positives and advance tool molecules and potential molecular glue candidates.
• Served as designated key personnel for laboratory safety and regulatory compliance, ensuring inspections to ensure zero violations.
• Delivered 130+ target protein–inhibitor X-ray structures, revealing key interactions and actionable design insights, documented in comprehensive reports.
• Supported structure-based drug design (SBDD) efforts across four projects in oncology, immuno-oncology, and neurological disorders, contributing to successful project progression to candidate selection.
• Designed functional surrogates for challenging proteins, enabling crystallization, structure determination, and advancement of SBDD/FBDD strategies.
• Conducted modeling studies to predict off-target interactions, leveraging cheminformatics to guide design of selective, potent molecules.
• Established a crystal-soak displacement workflow for high-throughput delivery of X-ray structures from HTS, DEL, and NMR fragment hits, facilitating rational design of selective and proprietary inhibitors by medicinal chemistry teams.
• Supported the protein sciences team by providing schematic protocols for the purification of difficult proteins, improving reproducibility and efficiency.
• Successfully mentored team members on crystallization strategies, achieving superior diffraction results and knowledge transfer across the team.
• Executed cloning, expression, purification, and X-ray structure determination of JNK2 kinase–inhibitor complexes (PDB: 7N8T, 8ELC).
• Mentored junior staff in gene cloning, protein purification, crystallization, and structure determination, fostering skill development and team capability.
• Authored intramural and extramural grant proposals, successfully contributing to laboratory funding and resource acquisition.
• Solved crystal structures of human KRAS GTPase bound to GDP, SML-8-73-1 inhibitor, and mutant forms [PDB IDs: 4LDJ, 4NMM, 4OBE, 4WA7, 4TQ9, 4QL3], providing insights into inhibitor binding and conformational dynamics.
• Conducted cloning, expression, purification, and characterization of TAK1 kinase inhibitors, including reversible (NG25, Takinib) and irreversible covalent compounds (SM1-71), using biophysical, biochemical, and cellular assays; pioneered meta-substituted pyrrolopyrimidine covalent TAK1 inhibitors [PDB IDs: 5V5N, 4O91, 5J9L, 5J8I, 5J7S, 5JH6, 5E7R, 5JK3].
• Designed and validated a selective irreversible Src kinase inhibitor (DGY-06-116), with structural validation [PDB IDs: 6ATE, 6E6E] and patent protection (U.S. Patent Application WO-2020257385-A1).
• Solved Glutathione S-transferase structure [PDB ID: 5J41], contributing to functional and structural insights for enzymatic studies.
• Analyzed polycyclic aromatic hydrocarbons (PAHs) in urban ambient air, coal tar combustion emissions, and diesel exhaust particles, assessing their genotoxic potential in human lung cells (A549 and BEAS-2B).
• Purified the ATPase domain of human topoisomerase-IIα and conducted biochemical inhibition studies with environmentally relevant quinone pollutants; quantified quinone-stabilized topoisomerase-IIα–DNA complexes using agarose-trapped DNA assays.
• Measured γ-H2AX levels in human lung cells as a marker of DNA double-strand breaks induced by quinone-mediated topoisomerase-IIα inhibition, linking environmental pollutants to cellular genotoxicity.
Crystallization and data collection: Co-crystallization methods, Crystal screening and freezing Laboratory automation and Robotics (Phoenix drop setter, Gryphon, Mosquito, Formulator, Rock Imager) Data collection (in-house and synchrotron beam lines), (XtaLAB MM003 XRD system, SAD/MAD experiments) Data processing (XDS, HKL2000, HKL3000, autoPROC)
Structure determination: Software suites-Global Phasing Buster, GRADE, Rhofit, CCP4, Coot, Phenix
Structure-based Drug Design (SBDD): XtalLead; Cheminformatics tools (Canvas, TIBCO Spotfire, ChemAxon, Virtual Screening, Chemical database mining, Spark library enumeration, QSAR, Chemical clustering of HTS, NMR hits
Modelling and visualization tools: (AlphaFold; Schrodinger Maestro suite: Pymol, Glide, SIFT, MD simulations; seeSAR, ChimeraX)
Gene editing and construct design: BLAST, Vector NTI, SnapGene, NCBI database, Uniprot, PCR, Cloning (Gibson, Ligation independent and Gateway)
Protein expression and purification: E coli, baculovirus-insect and mammalian cell expression, Purification using AKTA and Bio-RAD systems
Biochemical assays: SDS-PAGE; Western blot; mobility shift microfluidic assay, FRET assay, LanthaScreen assay, Kinetic parameters (kcat, Km; kinact, Ki)
Biophysical Assays: ITC; SPR; DSF
Cellular assays: Trapped in agarose DNA immunostaining (TARDIS) assay; The CellTiter-Glo Luminescent Cell Viability Assay (CTG)
Successful Academic Collaborations: Dana-Farber Cancer Institute, UT Southwestern Medical Center, Duke University
· Gurbani D, Du G, et al. Structure and Characterization of a Covalent Inhibitor of Src Kinase. Front. Mol. Biosci. May 2020, Volume 7, Article 81 | doi: 10.3389/fmolb.2020.00081
Designed a selective covalent Src kinase inhibitor [U.S. Patent Application WO-2020257385-A1] and determined its X-ray crystal structure.
· Du G, Rao S, Gurbani D*, et al. Structure-Based Design of a Potent and Selective Covalent Inhibitor for SRC Kinase That Targets a P-Loop Cysteine. J Med Chem. 2020 Jan 30. doi: 10.1021/acs.jmedchem.9b01502. *Equal Contribution.
Performed cloning, expression, purification, crystallization, and X-ray structure determination of Src kinase, alongside biophysical and biochemical characterization of covalent inhibitors.
· Totzke J., Gurbani D.*, et al. Takinib, an exquisitely selective inhibitor of TAK1, targets pro survival TNFα-dependent signaling, inducing apoptosis in rheumatoid arthritis and breast cancer models. Cell Chemical Biology 24 (2017), 1029–1039. *Equal Contribution.
Solved the X-ray crystal structure of Takinib bound to TAK1 kinase and elucidated the structure–activity relationship (SAR) of Takinib analogs.
· Hunter J. C., Gurbani D.,et al. In situ selectivity profiling and crystal structure of SML-8-73-1, an active site inhibitor of oncogenic K-Ras G12C, Proc Natl Acad Sci U S A, 111 (2014) 8895-8900. PMID: 24889603.
Crystallized and determined the X-ray structure of the covalent KRAS^G12C inhibitor SML-8-73-1.
· Tan L., Nomanbhoy T., Gurbani D., et al. Discovery of type II inhibitors of TGFbeta-activated kinase 1 (TAK1) and mitogen-activated protein kinase kinase kinase kinase 2 (MAP4K2), J Med Chem, 58 (2015) 183-196. PMID: 25075558.
Performed cloning, expression, purification, and crystallization of TAK1 kinase and determined multiple X-ray structures of inhibitor-bound complexes, supporting pharmacophore validation and rational design of Type II inhibitors.
Complete list (26+):
https://www.ncbi.nlm.nih.gov/myncbi/deepak.gurbani.1/bibliography/public
Delivered Invited Talk on 'Takinib as selective inhibitor of TAK1 kinase' at Cancer Center Biology Retreat on Friday, September 1, 2017, at Harold C. Simmons Comprehensive Cancer Center, UT Southwestern Medical Center.
Protein engineering, Crystallography, Structure based drug design, Assay development, Table Tennis, Badminton, Cooking
https://www.linkedin.com/in/deepak-gurbani-ph-d-53154634
https://scholar.google.com/citations?user=W7Px5isAAAAJ&hl=en&oi=ao
orcid.org/0000-0002-0982-7173