Vyacheslav (Slava) I. Kuznetsov, Ph.D
Madison,U.S.
Summary
Dynamic and results-driven scientist with over 10 years of diverse laboratory experience in enzymology, organic chemistry, biochemistry, and molecular biology. Expertise in the preparation and screening of drug candidates, synthesis of peptide reagents and arrays. Extensive experience in preparation characterization of enzymes, protein complexes and bio-sensors. Proven success in designing and executing multiple projects and collaborations, demonstrating a commitment to innovation and teamwork. Professional analytical and problem-solving skills, complemented by proficiency in multitasking, time management, and team building.
Overview
18
18
years of professional experience
Work History
Scientist II
Wisconsin Institute for Discovery, University of Wisconsin-Madison
04.2022 - Current
- synthetic preparation and validation of published Sirt6 modulators (activators and inhibitors);
- development of innovative approaches for large-scale preparation of recombinant and acetylated nucleosomes;
- design of novel large-scale PBP-MDCC purification of Pi-sensor;
- design and validation of novel high-throuput NUDIX/PBP-MDCC sirtuin plate-reader fluorescent assay;
Scientist Project Lead
Galilei Biosciences
01.2021 - Current
- chemical synthesis and kinetic evaluation of novel Sirt6 small molecule activators (SAR optimization of Cl5D lead);
- supervided collaboration between diverse research groups (Pharmaron (Crystallography), Immuto Scientific(PLIMB), O2H(SPR));
- analyzed experimental data to drive informed decision-making in project direction and execution;
Scientist I
Wisconsin Institute for Discovery, University of Wisconsin-Madison
12.2021 - 04.2022
- design of optimized HT-HPLC sirtuin assay;
- design and optimization of LC/MS-based sirtuin assay;
- design and validation of HT-fluorescent polarization (FP) sirtuin assay;
- design and validation of NanoLuc based sirtuin assay;
- cloning, optimized expression, purification and investigation of enzymatic properties of Sirt7;
Assistant Scientist
University of Wisconsin-Madison
08.2018 - 11.2021
Postdoctoral Research Associate
Research Group of John Denu, Department of Biomolecular Chemistry, School of Medicine and Public Health University of Wisconsin-Madison
01.2014 - 07.2018
- design and preparation of histone peptide arrays;
- cloning, expression, purification, and validation of HALO-tagged peptide arrays;
- Solid Phase Peptide Synthesis (peptide probes, peptide substrates, and peptide-arrays);
- isolation and LC/MS analysis of chromatin material from yeast, insect, and mammalian cell lines;
Postdoctoral Research Associate
Research Group of Alvan C Hengge Department of Chemistry and Biochemistry, Utah State University, Logan, UT, USA
06.2013 - 01.2014
- investigation of phosphotransfersase activity of select protein tyrosine phosphatases;
- training of new graduate students to use stop-flow, quench-flow, and steady-state enzyme kinetics;
- preparation and analysis of phosphopeptide arrays (384 well format);
Graduate Researcher/Teaching Assistant
Research Group of Alvan C Hengge, Department of Chemistry and Biochemistry Utah State University, Logan, UT, UT
08.2007 - 06.2013
Education
Ph.D. - Chemistry
Utah State University
Logan, UT, USA05.2013
B.Sc. - Pharmaceutical Technology
National University of Pharmacy
Kharkov, Ukraine05.2006
Skills
- Enzymology: assay development and mechanistic analysis of native (WT) and modified enzymes, protein-protein, and protein-DNA complexes Mechanistic investigation using Michaelis-Menten kinetics, kinetic isotope effects, steady-state, and pre-steady state kinetics (stopped-flow, quench-flow), mechanistic analysis of enzymatic modulators (inhibitors and activators)
- Protein crystallography: protein crystallization expertise: robotic (Mosquito Crystal) and manual optimization (siting/hanging drop), cryoprotection, collection of X-ray diffraction data, model refinement and validation Crystallized and deposited the following structures (PDPID): 4ERC, 4HAH, 4H43 Designed and supervised crystallization and refinement of several proprietary structures Software: HKL2000, HKL3000, COOT, PHENIX, CCP4, MOSFLM, PyMol
- Molecular Biology: DNA sequence optimization and cloning, recombinant protein production: codon-optimization, expression and co-expression in bacterial, insect, and mammalian hosts Chromatographic isolation under native and denaturing conditions, cryoprotection and lyophilization Global protein modifications: amber-codon suppression, chemical and enzymatic labeling, reductive methylation for protein crystallography Large scale production of RNA and DNA
- Tissue-culture: experienced at working in clean rooms and laminar flow hoods CRISPR-Cas9 genome modification/gene editing Production of recombinant RNPs, single colony isolation, chemical and viral protein and RNA cell transfection and transduction, subcellular fractionation and isolation of material
- Organic Chemistry: Skilled in conducting air/moisture/light-sensitive reactions; skilled in handling reactive /pyrophoric metals, metal hydrides and reagents; experienced in using Schlenk apparatus Manual and automated (Synergy) large-scale solid phase peptide synthesis (SPPS), high-throughput parallel peptide synthesis (Intavis), preparation of biotinylated peptide arrays and peptide-cellulose conjugates (CelluSpot)
- Compound purification/isolation: flash chromatography, prep-TLC, DCVC, Combi-Flash, HPLC (normal and reverse phase), re-crystallization, distillation, sublimation, precipitation and Soxhlet extractions
- Medicinal Chemistry: Design creative strategies for hit to lead (H2L) generation and lead optimization Analyze and interpret screening data, propose and implement SAR strategies
- Analytical skills Colorimetric, fluorimetric, and luminescent kinetic and binding assays Design and implementation of enzymatic coupled, high-throughput plate reader assays, immobilized peptide arrays, NMR, HPLC and LCMS analysis Design and analysis of high-throughput screening data and analysis of drug candidates
Languages
English
Native or Bilingual
Ukrainian
Native or Bilingual
Russian
Native or Bilingual
Timeline
Scientist II
Wisconsin Institute for Discovery, University of Wisconsin-Madison
04.2022 - Current
Scientist I
Wisconsin Institute for Discovery, University of Wisconsin-Madison
12.2021 - 04.2022
Scientist Project Lead
Galilei Biosciences
01.2021 - Current
Assistant Scientist
University of Wisconsin-Madison
08.2018 - 11.2021
Postdoctoral Research Associate
Research Group of John Denu, Department of Biomolecular Chemistry, School of Medicine and Public Health University of Wisconsin-Madison
01.2014 - 07.2018
Postdoctoral Research Associate
Research Group of Alvan C Hengge Department of Chemistry and Biochemistry, Utah State University, Logan, UT, USA
06.2013 - 01.2014
Graduate Researcher/Teaching Assistant
Research Group of Alvan C Hengge, Department of Chemistry and Biochemistry Utah State University, Logan, UT, UT
08.2007 - 06.2013
Ph.D. - Chemistry
Utah State University
B.Sc. - Pharmaceutical Technology
National University of Pharmacy
Teaching Experience
- Teaching assistant 2007 - 2013
- CHEM 1010 - General Chemistry
- CHEM 1210 - General Chemistry
- CHEM 2315 - Organic Chemistry
- CHEM 2325 - Organic Chemistry
- CHEM 5720 - Biochemistry
Selected Presentations at Professional Meetings
- 2017 “National Meeting of the American Society for Biochemistry and Molecular Biology, Chicago, IL New Method for Isolation of Native Yeast Nucleosomes - Development and Application
- 2012 “National Meeting of the American Society for Biochemistry and Molecular Biology, San Diego, CA.“High resolution structure of the phosphatase VHZ explains unexpected substrate specificity”
- 2012 Hansen Life Sciences Retreat 2012 Logan, UT
- 2010 National 239th Meeting of the ACS, San Francisco, CA. “Kinetic studies of the dual-specificity phosphatase VHZ” Division of Biological Chemistry, abstract #93.
- 2009 Intermountain Graduate Research Symposium, Logan, UT “Cloning, purification and preliminary kinetic analysis of VHZ”
List of Publications
In collaboration with: Prof. Anastassia Alexandrova, University of California; Prof. Sean Johnson, Utah State University; Prof. Catherine Fox, University of Wisconsin-Madison; Prof. Weiping Tang, University of Wisconsin-Madison; Prof. George N. Phillips, Rice University, St, Houston, TX; Prof. Ed Luk, Stony Brook University.
10. Comeros C, Pierrakeas L, Chan L, Chowdhury S, de Onis E, Kuznetsov, V.I, et al. (2025) H2A.Z deposition by the SWR complex is stimulated by polyadenine DNA sequences in nucleosomes. PLoS Biol 23(5).
9. Spencer A. Haws, Lillian J. Miller, Diego Rojas La Luz, Vyacheslav I. Kuznetsov, Raymond C. Trivel, Gheorghe Craciun, and John Denu (2023): "Intrinsic catalytic properties of histone H3 lysine-9 methyltransferases preserve monomethylation levels under low S-adenosylmethionine", Journal of Biological Chemistry, 299(7).
8. Vyacheslav I. Kuznetsov, Wallace H Liu, Mark A Klein, John M Denu (2022) “Potent Activation of NAD+-Dependent Deacetylase Sirt7 by Nucleosome Binding, ACS Chem Bio 17(8).
7. Wallace H Liu, Jie Zheng, Jessica L Feldman, Mark A Klein, Vyacheslav I Kuznetsov, Craig L Peterson, Patrick R Griffin, John Denu, “Multivalent interactions drive nucleosome binding and efficient chromatin deacetylation by SIRT6”, Nature communications 11 (1), 1-13.
6. Mark A Klein, Can Liu, Vyacheslav I Kuznetsov, John B Feltenberger, Weiping Tang, John M Denu. "Mechanism of activation for the sirtuin 6 protein deacylase", Journal of Biological Chemistry 295 (5), 1385-1399 Editor Highlighted.
5. Kuznetsov, V. I., Spencer A. Haws, Catherine Fox, John Denu "General method for rapid purification of native chromatin fragments", Journal of Biological Chemistry, 293 (31), 12271-12282
4. Su F, Wang J, Denu JM, Kuznetsov V. I., KE Stephens, R Papazyan, George N Phillips, John M Denu (2016) "Reader domain specificity and lysine demethylase-4 family function" Nature communications 7 (1), 1-15
3. Kuznetsov, V. I., Alexandrova, A. N., and Hengge, A. C. (2013) "New functional aspects of the atypical protein tyrosine phosphatase VHZ" Biochemistry 52 (45), 8012-8025
2. Kuznetsov, V. I., Hengge, A. C., and Johnson, S. J. (2012) "New Aspects of the Phosphatase VHZ Revealed by a High-Resolution Structure with Vanadate and Substrate Screening", Biochemistry 51 (49), 9869-9879
1. Kuznetsov, V. I., Alexandrova, A. N., and Hengge, A. C. (2012) Metavanadate at the Active Site of the Phosphatase VHZ, Journal of the American Chemical Society, 134, 14298-14301. Editor Highlighted