William Gutheil

Biography

My laboratory operates at the intersection of synthetic and analytical chemistry, biology, biophysics, enzymology, and drug discovery. My research program is focused on the discovery of novel antibacterial agents and synergistic antibacterial drug combinations, with particular emphasis on inhibitors of bacterial cell wall biosynthesis, their enzymatic targets, and mechanisms of antibiotic resistance. A core strength of my research is expertise in analytical biochemistry and mass spectrometry. I serve as Director of the UMKC Drug Discovery and Metabolomics LC-MS Shared Instrumentation Resource, where my laboratory has developed comprehensive LC- MS/MS methods for the quantitative analysis of all known cytoplasmic intermediates in bacterial cell wall biosynthesis, as well as DNA and RNA nucleosides and nucleotides. The recent acquisition of a high-resolution QToF mass spectrometer has further expanded these capabilities, enabling both high-sensitivity targeted analyses and untargeted metabolomics. These tools are being applied to define, at a systems level, how diverse antibiotics perturb cell wall biosynthesis in antibiotic-sensitive and antibiotic- resistant bacterial pathogens, and to elucidate mechanisms of action of novel antibacterial agents. In parallel, my laboratory has established a robust antibacterial library-screening and synergy-screening platform. Whole-library synergy screening of multiple high- value compound collections has identified numerous unexpected and mechanistically informative drug combinations. These studies demonstrate the strong potential of synergy screening both as a translational strategy for identifying clinically relevant antibacterial combinations and as a powerful probe of bacterial biochemistry and metabolic regulation.

Biological applications of LC-MS/MS technology. Dr. Gutheil established and oversees the Drug Discovery and Metabolomics LC-MS/MS shared instrumentation resource in the School of Pharmacy. This facility currently houses a Sciex 3200 QTrap instrument, an Agilent 6495c triple-quad instrument, and a Shimadzu 9030 QToF instrument. Faculty and graduate students in the School of Pharmacy and other UMKC units are provided training and hands on access to these instruments for use in small to medium sized molecule research. This facility is heavily used and very productive. Efforts to enhance this resource are ongoing.

Bibliography

1. Gutheil, W.G., Checkerboard synergistic data analysis using a Hill function-based approach. Next Research, 2026. 3: p. 101138.
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2. Mishra, N.R. and W.G. Gutheil, Stereoselective Amine-omics Using Heavy Atom Isotope Labeled L- and D-Marfey's Reagents. J Am Soc Mass Spectrom, 2024.Sharma, A.D. and W.G. Gutheil, Synergistic
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3. Combinations of FDA-Approved Drugs with Ceftobiprole against Methicillin-Resistant Staphylococcus aureus. Microbiol Spectr, 2023. 11(1): p. e0372622.
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4. Punchihewa, B.T., V. Minda, W.G. Gutheil, and M. Rafiee, Electrosynthesis and Microanalysis in Thin Layer: An Electrochemical Pipette for Rapid Electrolysis and Mechanistic Study of Electrochemical Reactions. Angew Chem Int Ed Engl, 2023. 62(44): p. e202312048.
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5. Mishra, N.R., A.D. Sharma, S. Gargvanshi, and W.G. Gutheil, Deconvolution of multichannel LC-MS/MS chromatograms of glucosamine-phosphates: Evidence of a GlmS regulatory difference between Staphylococcus aureus and Enterococcus faecium. Talanta Open, 2023. 8.
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6. Gargvanshi, S., G. Heravi, N.J. Ayon, and W.G. Gutheil, Screening the NCI diversity set V for anti-MRSA activity: cefoxitin synergy and LC-MS/MS confirmation of folate/thymidine biosynthesis inhibition. Microbiol Spectr, 2023. 11(6): p. e0054123.
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7. Gargvanshi, S. and W.G. Gutheil, Library Screening for Synergistic Combinations of FDA-Approved Drugs and Metabolites with Vancomycin against VanA-Type Vancomycin-Resistant Enterococcus faecium. Microbiol Spectr, 2022. 10(5): p. e0141222.
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8. Gargvanshi, S., H. Vemula, and W.G. Gutheil, Effect of Vancomycin on Cytoplasmic Peptidoglycan Intermediates and van Operon mRNA Levels in VanA-Type Vancomycin-Resistant Enterococcus faecium. J Bacteriol, 2021. 203(16): p. e0023021.
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9. Massa, S.M., A.D. Sharma, C. Siletti, Z. Tu, J.J. Godfrey, W.G. Gutheil, and T.N. Huynh, c-di-AMP Accumulation Impairs Muropeptide Synthesis in Listeria monocytogenes. J Bacteriol, 2020. 202(24).
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10. Dey, A.S., N.J. Ayon, C. Bhattacharya, W.G. Gutheil, and M. Mukherji, Positive/negative ion-switching-based LC-MS/MS method for quantification of cytosine derivatives produced by the TET-family 5-methylcytosine dioxygenases. Biol Methods Protoc, 2020. 5(1): p. bpaa019.
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11. Ayon, N.J., A.D. Sharma, and W.G. Gutheil, LC-MS/MS-Based Separation and Quantification of Marfey's Reagent Derivatized Proteinogenic Amino Acid DL-Stereoisomers. J Am Soc Mass Spectrom, 2019. 30(3): p. 448-58.
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12. Ayon, N.J. and W.G. Gutheil, Dimensionally Enhanced Antibacterial Library Screening. ACS Chem Biol, 2019. 14(12): p. 2887-94
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13. Kitase, Y., J.A. Vallejo, W. Gutheil, H. Vemula, K. Jahn, J. Yi, J. Zhou, M. Brotto, and L.F. Bonewald, beta-aminoisobutyric Acid, l-BAIBA, Is a Muscle-Derived Osteocyte Survival Factor. Cell Rep, 2018. 22(6): p. 1531-44.
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14. Gutheil, W.G., Derivation and numerical profile analysis of a hierarchically formulated microscopic model of hemoglobin oxygen binding. Biophys Chem, 2018. 241: p. 38-49.
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15. Bhattacharya, C., A.S. Dey, N.J. Ayon, W.G. Gutheil, and M. Mukherji, Efficient Purification and LC-MS/MS-based Assay Development for Ten-Eleven Translocation-2 5-Methylcytosine Dioxygenase. J Vis Exp, 2018(140).
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16. Vemula, H., Y. Kitase, N.J. Ayon, L. Bonewald, and W.G. Gutheil, Gaussian and linear deconvolution of LC-MS/MS chromatograms of the eight aminobutyric acid isomers. Anal Biochem, 2017. 516: p. 75-85.
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17. Vemula, H., N.J. Ayon, A. Burton, and W.G. Gutheil, Antibiotic Effects on Methicillin-Resistant Staphylococcus aureus Cytoplasmic Peptidoglycan Intermediate Levels and Evidence for Potential Metabolite Level Regulatory Loops. Antimicrob Agents Chemother, 2017. 61(6).
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18. Jaiswal, M., S. Bhar, H. Vemula, S. Prakash, V.K.C. Ponnaluri, W.G. Gutheil, and M. Mukherji, Convenient expression, purification and quantitative liquid chromatography-tandem mass spectrometry-based analysis of TET2 5-methylcytosine demethylase. Protein Expr Purif, 2017. 132: p. 143-51.
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19. Vemula, H., N.J. Ayon, and W.G. Gutheil, Cytoplasmic peptidoglycan intermediate levels in Staphylococcus aureus. Biochimie, 2016. 121: p. 72-8.
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20. Rangarajan, P., et al., Crocetinic acid inhibits hedgehog signaling to inhibit pancreatic cancer stem cells. Oncotarget, 2015. 6(29): p. 27661-73.
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21. Vemula, H., S. Bobba, S. Putty, J.E. Barbara, and W.G. Gutheil, Ion-pairing liquid chromatography-tandem mass spectrometry-based quantification of uridine diphosphate-linked intermediates in the Staphylococcus aureus cell wall biosynthesis pathway. Anal Biochem, 2014. 465: p. 12-9.
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22. Putty, S., H. Vemula, S. Bobba, and W.G. Gutheil, A liquid chromatography-tandem mass spectrometry assay for D-Ala-D-Lac: a key intermediate for vancomycin resistance in vancomycin-resistant enterococci. Anal Biochem, 2013. 442(2): p. 166-71.
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23. Paulillo, L.C., C. Mo, J. Isaacson, L. Lessa, E. Lopes, S. Romero-Suarez, L. Brotto, E. Abreu, W. Gutheil, and M. Brotto, Jatropha curcas: from biodiesel generation to medicinal applications. Recent Pat Biotechnol, 2012. 6(3): p. 192-9.
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24. Gutheil, W.G., G. Reed, A. Ray, S. Anant, and A. Dhar, Crocetin: an agent derived from saffron for prevention and therapy for cancer. Curr Pharm Biotechnol, 2012. 13(1): p. 173-9.
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25. Bobba, S., G.E. Resch, and W.G. Gutheil, A liquid chromatography-tandem mass spectrometry assay for detection and quantitation of the dipeptide Gly-Gln in rat brain. Anal Biochem, 2012. 425(2): p. 145-50.
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26. Putty, S., A. Rai, D. Jamindar, P. Pagano, C.L. Quinn, T. Mima, H.P. Schweizer, and W.G. Gutheil, Characterization of d-boroAla as a novel broad-spectrum antibacterial agent targeting d-Ala-d-Ala ligase. Chem Biol Drug Des, 2011. 78(5): p. 757-63.
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27. Bobba, S., V.K. Ponnaluri, M. Mukherji, and W.G. Gutheil, Microtiter plate-based assay for inhibitors of penicillin-binding protein 2a from methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother, 2011. 55(6): p. 2783-7.
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28. Bobba, S. and W.G. Gutheil, Multivariate geometrical analysis of catalytic residues in the penicillin-binding proteins. Int J Biochem Cell Biol, 2011. 43(10): p. 1490-9.
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29. Stefanova, M., S. Bobba, and W.G. Gutheil, A microtiter plate-based beta-lactam binding assay for inhibitors of high-molecular-mass penicillin-binding proteins. Anal Biochem, 2010. 396(1): p. 164-6.
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30. Singh, B., S.M. Mense, N.K. Bhat, S. Putty, W.A. Guthiel, F. Remotti, and H.K. Bhat, Dietary quercetin exacerbates the development of estrogen-induced breast tumors in female ACI rats. Toxicol Appl Pharmacol, 2010. 247(2): p. 83-90.
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31. Jamindar, D. and W.G. Gutheil, A liquid chromatography-tandem mass spectrometry assay for Marfey's derivatives of L-Ala, D-Ala, and D-Ala-D-Ala: application to the in vivo confirmation of alanine racemase as the target of cycloserine in Escherichia coli. Anal Biochem, 2010. 396(1): p. 1-7.
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32. Ponnaluri, V.K., D.T. Vavilala, S. Putty, W.G. Gutheil, and M. Mukherji, Identification of non-histone substrates for JMJD2A-C histone demethylases. Biochem Biophys Res Commun, 2009. 390(2): p. 280-4.
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33. Peddi, S., R.A. Nicholas, and W.G. Gutheil, Neisseria gonorrhoeae penicillin-binding protein 3 demonstrates a pronounced preference for N(epsilon)-acylated substrates. Biochemistry, 2009. 48(24): p. 5731-7.
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34. Rai, A. and W.G. Gutheil, A Dde resin based strategy for inverse solid-phase synthesis of amino terminated peptides, peptide mimetics and protected peptide intermediates. J Pept Sci, 2005. 11(2): p. 69-73.
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35. Nicola, G., S. Peddi, M. Stefanova, R.A. Nicholas, W.G. Gutheil, and C. Davies, Crystal structure of Escherichia coli penicillin-binding protein 5 bound to a tripeptide boronic acid inhibitor: a role for Ser-110 in deacylation. Biochemistry, 2005. 44(23): p. 8207-17.
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36. Stefanova, M.E., J. Tomberg, C. Davies, R.A. Nicholas, and W.G. Gutheil, Overexpression and enzymatic characterization of Neisseria gonorrhoeae penicillin-binding protein 4. Eur J Biochem, 2004. 271(1): p. 23-32.
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37. Sasubilli, R. and W.G. Gutheil, General inverse solid-phase synthesis method for C-terminally modified peptide mimetics. J Comb Chem, 2004. 6(6): p. 911-5.
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38. Stefanova, M.E., J. Tomberg, M. Olesky, J.V. Holtje, W.G. Gutheil, and R.A. Nicholas, Neisseria gonorrhoeae penicillin-binding protein 3 exhibits exceptionally high carboxypeptidase and beta-lactam binding activities. Biochemistry, 2003. 42(49): p. 14614-25.
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39. Pechenov, A., M.E. Stefanova, R.A. Nicholas, S. Peddi, and W.G. Gutheil, Potential transition state analogue inhibitors for the penicillin-binding proteins. Biochemistry, 2003. 42(2): p. 579-88.
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40. Stefanova, M.E., C. Davies, R.A. Nicholas, and W.G. Gutheil, pH, inhibitor, and substrate specificity studies on Escherichia coli penicillin-binding protein 5. Biochim Biophys Acta, 2002. 1597(2): p. 292-300.
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41. Gutheil, W.G. and Q. Xu, N-to-C solid-phase peptide and peptide trifluoromethylketone synthesis using amino acid tert-butyl esters. Chem Pharm Bull (Tokyo), 2002. 50(5): p. 688-91.
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42. Gutheil, W.G., M.E. Stefanova, and R.A. Nicholas, Fluorescent coupled enzyme assays for D-alanine: application to penicillin-binding protein and vancomycin activity assays. Anal Biochem, 2000. 287(2): p. 196-202.
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43. Gutheil, W.G., Application of hierarchical thermodynamic interactions to the protonation equilibria of organic polyprotic acids. Biophys Chem, 2000. 88(1-3): p. 119-26.
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44. Gutheil, W.G., A simple chemical example of hierarchical thermodynamic interactions: the protonation equilibria of inorganic polyprotic acids. Biophys Chem, 2000. 88(1-3): p. 35-45.
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45. Gutheil, W.G., Statistical analysis of data pertaining to complex state systems by stepwise regression with reformulated parameters; application to spectroscopically monitored hemoglobin oxygen binding data. Biophys Chem, 1998. 70(3): p. 185-202.
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46. Gutheil, W.G., A sensitive equilibrium-based assay for D-lactate using D-lactate dehydrogenase: application to penicillin-binding protein/DD-carboxypeptidase activity assays. Anal Biochem, 1998. 259(1): p. 62-7.
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47. Gutheil, W.G., E. Kasimoglu, and P.C. Nicholson, Induction of glutathione-dependent formaldehyde dehydrogenase activity in Escherichia coli and Hemophilus influenza. Biochem Biophys Res Commun, 1997. 238(3): p. 693-6.
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48. Tsilikounas, E., T. Rao, W.G. Gutheil, and W.W. Bachovchin, 15N and 1H NMR spectroscopy of the catalytic histidine in chloromethyl ketone-inhibited complexes of serine proteases. Biochemistry, 1996. 35(7): p. 2437-44.
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49. Snow, R.J., W.W. Bachovchin, R.W. Barton, S.J. Campbell, S.J. Coutts, D.M. Freeman, W.G. Gutheil, T.A. Kelly, and C.A. Kennedy, Studies on Proline Boronic Acid Dipeptide Inhibitors of Dipeptidyl Peptidase IV: Identification of a Cyclic Species Containing a B-N Bond. Journal of the American Chemical Society, 1994. 116(24): p. 10860-9.
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50. Gutheil, W.G., M. Subramanyam, G.R. Flentke, D.G. Sanford, E. Munoz, B.T. Huber, and W.W. Bachovchin, Human immunodeficiency virus 1 Tat binds to dipeptidyl aminopeptidase IV (CD26): a possible mechanism for Tat's immunosuppressive activity. Proc Natl Acad Sci U S A, 1994. 91(14): p. 6594-8.
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51. Gutheil, W.G., C.A. Kettner, and W.W. Bachovchin, Kinlsq: a program for fitting kinetics data with numerically integrated rate equations and its application to the analysis of slow, tight-binding inhibition data. Anal Biochem, 1994. 223(1): p. 13-20.
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52. Gutheil, W.G., Reformulation of thermodynamic systems with aggregation and theoretical methods for the analysis of ligand binding in proteins with monomer-multimer equilibria. Biophys Chem, 1994. 52(1): p. 83-95.
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53. Broder, C.C., O. Nussbaum, W.G. Gutheil, W.W. Bachovchin, and E.A. Berger, CD26 antigen and HIV fusion? Science, 1994. 264(5162): p. 1156-9; author reply 62-5.
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54. Farr-Jones, S., W.Y.L. Wong, W.G. Gutheil, and W.W. Bachovchin, Direct observation of the tautomeric forms of histidine in nitrogen-15 NMR spectra at low temperatures. Comments on intramolecular hydrogen bonding and on tautomeric equilibrium constants. Journal of the American Chemical Society, 1993. 115(15): p. 6813-9.
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55. Subramanyam, M., W.G. Gutheil, W.W. Bachovchin, and B.T. Huber, Mechanism of HIV-1 Tat induced inhibition of antigen-specific T cell responsiveness. J Immunol, 1993. 150(6): p. 2544-53.
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56. Gutheil, W.G. and W.W. Bachovchin, Separation of L-Pro-DL-boroPro into its component diastereomers and kinetic analysis of their inhibition of dipeptidyl peptidase IV. A new method for the analysis of slow, tight-binding inhibition. Biochemistry, 1993. 32(34): p. 8723-31.
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57. Dahl, N.K., W.G. Gutheil, and L. Liscum, Abnormal regulation of low density lipoprotein-sensitive events in a cholesterol transport mutant. J Biol Chem, 1993. 268(23): p. 16979-86.
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58. Gutheil, W.G. and C.E. McKenna, Unique and independent parameters (UIP) formulation for thermodynamic models of complex protein-ligand systems. Biophys Chem, 1992. 45(2): p. 171-9.
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59. Gutheil, W.G., B. Holmquist, and B.L. Vallee, Purification, characterization, and partial sequence of the glutathione-dependent formaldehyde dehydrogenase from Escherichia coli: a class III alcohol dehydrogenase. Biochemistry, 1992. 31(2): p. 475-81.
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60. Gutheil, W.G., Thermodynamic model of cooperativity in a dimeric protein: unique and independent parameters formulation. Biophys Chem, 1992. 45(2): p. 181-91.
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61. McKenna, C.E., W.G. Gutheil, and W. Song, A method for preparing analytically pure sodium dithionite. Dithionite quality and observed nitrogenase-specific activities. Biochim Biophys Acta, 1991. 1075(1): p. 109-17.
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