Research Interests

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We are developing biophysical methods using mass spectrometry to understand protein interactions. One method is PLIMSTEX (Protein Ligand Interaction by Mass Spectrometry, Titration and H/D Exchange). The figure shows the titration of fatty acid binding protein (structure on left) with the fatty-acid ligand, potassium oleate, to give a complex (on right) that now contains the fatty-acid anion. The y-axis data are mass spectrometric molecular weight measurements, and the solid line is computed from our model that gives the equilibrium constant and protection factor for the protein-ligand interaction.

 

A more specific approach for footprinting is to map proteins with reactions of OH radicals. In FPOP (fast photochemical oxidation of proteins), we “paint” certain surface-accessible amino-acid residues by reacting the protein in a flow tube (see figure below) with radicals generated from H₂O₂ by a pulsed KrF laser (248 nm). With appropriate scavengers, the reaction time is remarkable short (~ 1 microsec). Certain amino acids react with the radicals to become oxidized (depicted as turning from “green” to “yellow”--see figure below). PLIMSTEX and FPOP may be useful for determining protein-protein interfaces, affinities for tiny amounts of proteins in biophysics and drug discovery.

Another goal is development of Fourier transform MS. Of interest are accurate mass measurement and new trap designs (see figure below) designed to incorporate electric field compensation. Two instruments equipped with 7-tesla magnets, and soon a 12-tesla instrument for top-down proteomics are available for fundamental and applied research.

Selected Publications

Microsecond Time-Scale Hydroxyl Radical Profiling of Solvent-Accessible Protein Residues, David M. Hambly and Michael L. Gross, chapter in Compehensive Analytical Chemistry, Volume 52, 151-177 (2009).

Combined Charged Residue-Field Emission Model of Macromolecular Electrospray Ionization, C.J. Hogan Jr, J.A. Carroll, H.W. Rohrs, P. Biswas, and Michael L. Gross, Anal. Chem., 81-369-377, (2009).

The Balance of Benefical and Deleterious health Effects of Quinoes:  A Case Study of the Chemical Properties of Genistein and Estrone Quinones, Qiang Zhang, Tingting Tu, Andre d’Avignon, and Michael L. Gross, J. Am. Chem. Soc., 131, 1067-1076, (2009).

2-Nitrophenyl aryl sulfides undergo both intramolecular and electrospray-induced intermolecular oxidation of sulfur:  An experimental and theoretical case study, Joseph T. Moolayil, M. George, Daryl Giblin, Michael L. Gross, Int J. Mass Spectrom., 283, 222-228, (2009).

Isolation and Identification of Two Novel SDS-Resistant Secreted Chitinases from Aeromonas schubertii, Chao-Lin, Chia-Rui Shen, Fong-Fu Hsu, Jeen-Kuan Chen, Pei-Tzu Wu, Shang-Hsin Guo, Wen-Chien Lee, Feng-Wei Yu, Zachary B. Mackey, John Turk, and Michael L. Gross, Biotechnol. Prog., 25, 1, 124-131, (2009).

Method Revealing Bacterial Cell-Wall Architecture by Time-Dependent Isotope Labeling and Quantitative Liquid Chromatography/Mass Spectrometry, Gary J. Patti, Jiawei Chen, and Michael L. Gross, Anal. Chem., 81, 2437-2455, (2009).

Gas-Phase Nazarov Cyclization of Protonated 2-Methoxy and 2-Hydroxychalcone:  An Example of Intramolecular Proton-Transport Catalysis, Mathai George, Valarkottu S. Sebastian, Putluri Nagi Reddy, Ragampeta Srinivas, Daryl Giblin, and Michael L. Gross, J. Am. Soc. Mass Spectrom,  20, 805-818, (2009).

Membrane orientation of the FMO antenna protein from Chlorobaculum tepidum as determined by mass spectrometry-based footprinting, Jianzhong. Wen, Hao Zhang, Michael L. Gross, and Robert E. Blankenship, Proceedings of the National Academy of Sciences of the United States of America, 106, 15, 6134-6139, (2009).

Radical Cation/Radical Reactions:  A Fourier Transform Ion Cyclotron Resonance Study of Allyl Radical Reacting with Aromatic Radical Cations, Amber L. Russell, Henry W. Rohrs, David Read, Daryl .E. Giblin, Peter P. Gaspar, Michael L. Gross, Int J. Mass Spectrom., 287, 8-15, (2009).

An Endogenous Peptide Positively Selects and Augments the Activation and Survival of Peripheral CD4⁺ T cells, Wan-Lin Lo, Nathan J. Felix, James J. Walters, Henry Rohrs, Michael L. Gross, Paul M. Allen, Nature Immunology, 10, 1155-1161, (2009).

The Gramicidin Dimer Shows Both EX1 and EX2 Mechanisms of H/D Exchange, Raghu K. Chitta, Don L. Rempel, and Michael L. Gross, J. Am. Soc. Mass Spectrom,  20, 1813-1820, (2009).

Photocrosslinking of Human Telomeric G-quadruplex Loops by anti Cyclobutane Thymine Dimer Formation, Dian G.T. Su, Huafeng Fang, Michael L. Gross, John-Stephen A.Taylor, Proceedings of the National Academy of Sciences of the United States of America, 106, 12861-12866, (2009).

Miniaturizing Sample Spots for Matrix-assisted Laser Desorption/ionization Mass Spectrometry, Tingting Tu and Michael L. Gross, Trends in Analytical Chemistry, 28, 833-841, (2009). 

Fast Photochemical Oxidation of Protein Footprints Faster than Protein Unfolding, Brian C. Gau, Joshua S. Sharp, Don L. Rempel, and Michael L. Gross, Anal. Chem., 81, 6563-6571, (2009).

Cold Chemical Oxidation of Proteins, David M. Hambly and Michael L. Gross, Anal. Chem., 81, 7235-7242, (2009).

Organization of Lower Rim O-Alkylated p-Phosphonic Acid Calix[4]arenes, Thomas E. Clark, Mohamed Makha, Alexandre N. Sobolev, Dian Su, Henry Rohrs, Michael L. Gross, and Colin L. Raston, Crystal Growth & Design, 9, 3575-3580, (2009).

A Single Peptide–MHC Complex Positively Selects a Diverse and Specific CD8 T Cell Repertoire, Baomei Wang, Tina M. Primeau, Nancy Myers, Henry W. Rohrs, Michael L. Gross, Lonnie Lybarger, Ted H. Hansen, Janet M. Connolly, Science, 326, 871-874, (2009).

Online, High-Pressure Digestion System for Protein Characterization by Hydrogen/Deuterium Exchange and Mass Spectrometry, Lisa M. Jones, Hao Zhang, Ilan Vidavsky, and Michael L. Gross, Anal. Chem., 82, 1171-1174, (2010).

A Tuning Method for Electrically Compensated Ion Cyclotron Resonance Mass Spectrometer Traps, Adam M. Brustkern, Don L. Rempel, and Michael L. Gross, J. Am. Soc. Mass Spectrom,  21, 451-454 (2010).

A New Photoproduct of 5-Methylcytosine and Adenine Characterized by HPLC and Mass Spectrometry, Dian G. T. Su, John-Stephen A. Taylor, and Michael L. Gross, Chem. Res. Toxicol,  23, 474-479 (2010)

 Protein-Peptide Affinity Determination Using an H/D Exchange Dilution Strategy: Application to Antigen-Antibody Interactions, Tingting Tu, T., M. Drǎguşanu, B.-A. Petre, Don L. Rempel, Michael Przybylski, and Michael L. Gross, J. Am. Soc. Mass Spectrom,  21 1660-1667 (2010).

 Sulfate radical anion as a new reagent for fast photochemical oxidation of proteins, Brian C. Gau, B.C., Hao Chen, Y. Zhang, and Michael L. Gross, Anal. Chem. 82 7821-7827 (2010).

 Ion behavior in an electrically compensated ion cyclotron resonance trap, Adam M. Brustkern, Don L. Rempel, and Michael L. Gross, Int. J. Mass Spectrom., in press.

 Radio Frequency-Only-Mode Event and Trap Compensation in Penning Fourier Transform Mass Spectrometry, A. M. Brustkern, D. L. Rempel, and Michael L. Gross, in Practical Aspects of Trapped Ion Mass Spectrometry, Vol. IV, Theory and Instrumentation, R.E. March and J. F. J. Todd, CRC Press, 433-468 (2010).

 Native Electrospray and Electron-Capture Dissociation in FTICR Mass Spectrometry Provide Top-Down Sequencing of a Protein Component in an Intact Protein Assembly, Hao Zhang, Weidong Cui, Jianzhong Wen, Robert E. Blankenship, Michael L. Gross, J. Am. Soc. Mass Spectrom. in press (2010).

Hydrophobic Peptides Affect Binding of Calmodulin and Ca²⁺ as Explored by H/D Amide Exchange and Mass Spectrometry, Justin B. Sperry, Richard Y-C. Huang, Mei M. Zhu, Don L. Rempel, and Michael L. Gross, Int. J. Mass Spectrom. in press (2010)

Improved mass spectrometric characterization of protein glycosylation reveals unusual glycosylation of maize-derived bovine trypsin, Hao Zhang, Richard Y-C. Huang, Pegah R. Jalili, Janet W. Irungu, Gordon R. Nicol, Kevin B. Ray, Henry W. Rohrs, Michael L. Gross, Anal. Chem. in press.

T-jump and Fast Photochemical Oxidation Probe Sub Millisecond Protein Folding, Jiawei Chen, Don L. Rempel, Michael L. Gross, J. Am. Chem. Soc. 132, 15502–15504 (2010).

 Structural model and spectroscopic characteristics of the FMO antenna protein from the aerobic chlorophototroph, Candidatus Chloracidobacterium thermophilum. Jianzhong Wen, Yusuke Tsukatani, Weidong Cui, Hao Zhang, Michael L. Gross, Donald A. Bryant, Robert E. Blankenship Biochim Biophys Acta  1807 157–164 (2011)

 Interactions of APE1 with a redox inhibitor: Evidence for an alternate conformation of the enzyme, Dian Su, Sarah Delaplane, Meihua Luo,  Don L. Rempel, Bich Vu,  Mark R. Kelley, Michael L. Gross, and Millie M. Georgiadis, Biochem. in press.

 Mass Spectrometry and Fast Photochemical Oxidation of Proteins (FPOP) for Comparing Structures of Protein/Ligand Complexes: The Calmodulin-peptide Model System, Hao Zhang, Brian C. Gau, Lisa M. Jones, Ilan Vidavsky, Michael L. Gross, Anal. Chem., in press.

Improved mass spectrometric characterization of protein glycosylation reveals unusual glycosylation of maize-derived bovine trypsin, Hao Zhang, Richard Y-C. Huang, Pegah R. Jalili, Janet W. Irungu, Gordon R. Nicol, Kevin B. Ray, Henry W. Rohrs, Michael L. Gross, Anal. Chem. 82, 10095-10101 (2010).

T-jump and Fast Photochemical Oxidation Probe Sub Millisecond Protein Folding, Jiawei Chen, Don L. Rempel, Michael L. Gross, J. Am. Chem. Soc. 132, 15502–15504 (2010).

Layered Calcium Structures of p-Phosphonic Acid-methyl-calix[6]arene, Thomas E. Clark, Adam Martin, Mohamed Makha, Alexandre N. Sobolev, Dian Su, Henry W. Rohrs, Michael L. Gross, and Colin L. Raston, Cryst. Growth Design, 10, 3211-3217 (2010)

Ion behavior in an electrically compensated ion cyclotron resonance trap, Adam M. Brustkern, Don L. Rempel, and Michael L. Gross, Int. J. Mass Spectrom., 300, 143-148 (2011).

Hydrophobic Peptides Affect Binding of Calmodulin and Ca²⁺ as Explored by H/D Amide Exchange and Mass Spectrometry, Justin B. Sperry, Richard Y-C. Huang, Mei M. Zhu, Don L. Rempel, and Michael L. Gross, Int. J. Mass Spectrom. 302, 85–92 (2011)

Structural model and spectroscopic characteristics of the FMO antenna protein from the aerobic chlorophototroph, Candidatus Chloracidobacterium thermophilum. Jianzhong Wen, Yusuke Tsukatani, Weidong Cui, Hao Zhang, Michael L. Gross, Donald A. Bryant, Robert E. Blankenship, Biochim Biophys Acta, 1807, 157–164 (2011)

Mass Spectrometry and Fast Photochemical Oxidation of Proteins (FPOP) for Comparing Structures of Protein/Ligand Complexes: The Calmodulin-peptide Model System, Hao Zhang, Brian C. Gau, Lisa M. Jones, Ilan Vidavsky, Michael L. Gross, Anal. Chem., 83, 311–318 (2011).

Interactions of APE1 with a redox inhibitor: Evidence for an alternate conformation of the enzyme, Dian Su, Sarah Delaplane, Meihua Luo, Don L. Rempel, Bich Vu, Mark R. Kelley, Michael L. Gross, and Millie M. Georgiadis, Biochem. 50, 82-92 (2011).

 Functional Analysis of Novel Analogs of E3330 that Block the Redox Signaling Activity of the Multifunctional AP Endonuclease/redox Signaling Enzyme APE1/Ref-1, Mark R. Kelley, Meihua Luo, April Reed, Dian Su, Sarah Delaplane, Richard Borch, Rodney L Nyland II, Michael L Gross, Millie Georgiadis, Antioxidants & Redox Signaling, 14, 1387-1401 (2011).

 Native Electrospray Mass Spectrometry Reveals the Nature and Stoichiometry of Pigments in the FMO Photosynthetic Antenna Protein, Jianzhong Wen, Hao Zhang, Michael L. Gross, and Robert E. Blankenship, Biochem., 50, 3502-3511 (2011).

 Mapping the protein-protein interface between a toxin and its cognate antitoxin from the bacterial pathogen Streptococcus pyogenes, Justin B. Sperry, Craig L. Smith, CL, Michael G. Caparon, Tom Ellenberger, Michael L. Gross, ML, Biochem., 50, 4038-4045 (2011).

 Carboxyl-Group Footprinting Maps the Dimerization Interface and Phosphorylation-induced Conformational Changes of a Membrane-associated Tyrosine Kinase, Hao Zhang, Wei Shen, Don Rempel, John Monsey, Ilan Vidavsky, Michael L. Gross, and Ron Bose, Molecular Cellular Proteomics, 2011 10(6), 1-16 M110.005678.  PubMed PMID: 21422241; PubMed Central PMCID: PMC3108836. (2011).

Structural determinant of chemical reactivity and potential health effects of quinones from natural products, Tingting Tu, Daryl Giblin, and Michael L. Gross, Chem. Research Toxicology, 24, 1527-1539 (2011)

HD Exchange and PLIMSTEX Determine the Affinities and Order of Binding of Ca2+ with Troponin C, Richard Y-C. Huang, Don L. Rempel, and Michael L. Gross, Biochem., 50, 5426–5435 (2011).

Native electrospray and electron-capture dissociation FTICR mass spectrometry for top-down studies of protein assemblies, Hao Zhang, Weidong Cui, Jianzhong Wen, Robert E. Blankenship, Michael L. Gross, Anal. Chem., 83, 5598-5606 (2011).

 Fast photochemical oxidation of proteins for epitope mapping, Lisa M. Jones, Justin B. Sperry, James A. Carroll,  Michael L. Gross, Anal. Chem., 83, 7657-7661 (2011)

Top-down Mass Spectrometry: Recent Developments, Applications, and Perspectives, Weidong Cui, Henry W. Rohrs and Michael L. Gross, Analyst, 136, 3854–3864 (2011).

Deprotonated N-(2,4-dinitrophenyl)amino acids undergo cyclization in solution and the gas phase, M. George, V. Ramesh, R. Srinivas, Daryl Giblin, Michael L. Gross, Intl. J. Mass Spectrom. 306, 232– 240 (2011)

Mass spectrometry-based protein footprinting characterizes the structures of oligomeric apolipoprotein E2, E3, and E4, Brian Gau, Kanchan Garai,  Carl Frieden, and Michael L. Gross, Biochemistry, 50, 8117-8126 (2011)

Hydrogen/Deuterium Exchange and Electron-Transfer Dissociation Mass Spectrometry Determine the Interface and Dynamics of Apolipoprotein E Oligomerization, Richard Y-C. Huang, Kanchan Garai, Carl Frieden, and Michael L. Gross, Biochemistry, 50, 9273–9282 (2011)

Psb27, a transiently associated protein, binds to the chlorophyll binding protein CP43 in photosystem II assembly intermediates, Haijun Liu, Richard Y.-C. Huang, Jiawei Chen, Michael L. Gross, and Himadri B. Pakrasi, Proceedings National Academy Sciences (USA), 108, 18536-18541 (2011).

S-Glutathionylation of cysteine 99 in the APE1 protein impairs abasic endonuclease activity, Yun-Jeong Kim, Daemyung Kim, Jennifer L Illuzzi, Sarah Delaplane; Dian Su; Michel Bernier, Michael L Gross; Millie M Georgiadis; David M. Wilson, III, Journal of Molecular Biology, 414, 313-326 (2011).

Chemical Footprinting for Determining Protein Properties and Interactions, by Sandra A. Kerfoot and Michael L. Gross, in Protein and Peptide Mass Spectrometry in Drug Discovery ed. by M.L. Gross, G. Chen, and B. Pramanik, John Wiley and Sons, Inc, Hoboken, NJ, Chapter 13, 175-211(2012)

Autoproteolytic Fragments Are Intermediates in the Oligomerization/Aggregation of the Parkinson's Disease Protein Alpha-Synuclein as Revealed by Ion Mobility Mass Spectrometry, C Vlad, K Lindner, C Karreman, S Schildknecht, M Leist M, N Tomczyk, J Rontree, J Langridge, K Danzer, T Ciossek, Alina Petre, Michael L Gross, B Hengerer, Michael Przybylski, ChemBioChem, 12, 2740-27444 (2011). 

Awards

2011, Mass Spectrometry Society of Japan, Honorary Lifetime Membership
2008, Bruker Daltonics Lectureship, University of Konstanz, Germany
2008, Freiser Lectureship, Purdue University
2006, J.J. Thomson Medal for Service to Int'l MS, Foundation for International Mass Spectrometry
2005, Excellence in Mentoring Award, Washington University Graduate Student Senate
2005, New Jersey Distinguished Lectureship, NJ Section ACS Mass Spectrometry Discussion Group
2004, Outstanding Mentor Award, Washington University Graduate Student Senate
2002, Midwest Award for Achievements in Chemistry, American Chemical Society
2002, Excellence in Mentoring Award, Washington University Graduate Student Senate
2001, Watkins Visiting Professor, Witchita State University
2001, Outstanding Mentor Award, Washington University Graduate Student Senate
2000, Frontiers in Chemistry Lectureship, Wayne State University
2000, Plenary Lecture, Vienna Mass Spectrometry Colloquium
2000, Plenary Lecture, EnviroAnalysis 2000
1999, Field and Franklin Award, American Chemical Society
1998, Plenary Lecture, Assoc. Official Anal. Chemists' Int'l Meeting
1998, Plenary Lecture, HPLC International Symposium
1997, Plenary Lecture, Colloquium Spectroscopicum Internationale
1997, Plenary Lecture, Igler MS Tage, Innsbruck
1996, Guest Faculty, NATO School on Biol. Mass Spectrometry
1995, Pioneers in Chemistry Series, Texas A&M University
1995, Plenary Lecture, Western Biotechnology Conference, San Diego
1995, Plenary Lecture, FDA Mass Spectrom. Symposium, Washington DC
1994, Plenary Lecture, Australia/New Zealand MS Conference, Melbourne
1993, Guest Professor, NATO Adv. Study Inst. Biol. MS, Ischia, Italy
1993, Plenary Lecture, Colloquium Spectroscopium Internationale
1993, Plenary Lecture, Taiwan Int'l Chemistry Conference
1993, Plenary Lecture, Indian Society for Mass Spectrometry
1993, Lifetime Member, Indian Society for Mass Spectrometry
1993, Kraft Lectureship, Indiana University
1992, Plenary Lecture, European Tandem Mass Spectrom. Conf.
1992, Recognized as 50 Most Cited Chemists, Institute for Scientific Information, 1984-1991
1991, Plenary Lecture, Australian Symposium on Analytical Chem.
1991, Plenary Lecture, Elsevier Anal. Advances for the 90s Symposium
1990, Van't Hoff Visiting Professor, Int'l Graduate School, University of Amsterdam
1990, Guest Pfrofessor, NATO Adv. Study Inst. in Biol. Mass Spectrometry
1988, Albright & Wilson Visiting Professor, University of Warwick
1988, Plenary Lecture, American Society for Mass Spectrometry
1987, Pioneer Award, Commonwealth of Mass "In Search for the Health Consequences of Dioxin in Our Environment."
1987, Hertel Lectureship, University of Tennessee
1987, Alan Maccoll Plenary Lectureship, British Royal Society for Mass Spectrometry
1987, Plenary Lecture, British Royal Society "Spectroscopy Across Spectrum"
1986, J.C. Karcher Lectureship, University of Oklahoma
1986, Plenary Lectureship, Versailles International Workshop on Ion-Molecule Rxs.
1984, Distinguished Lecture Series, Indiana University
1984, Plenary Lecture, Swiss/French Mass Spectrometry Conference
1984, Third Cycle Lectures, Eidegenössische Technische Hochschule Lausanne
1978, Foundation Medal for Distinguished Teaching, University of Nebraska

Courses

Chemistry 112, General Chemistry
Chemistry 152, General Chemistry Laboratory
Chemistry 550, Advanced Topics in Mass Spectrometry