DBBS

Our principal research goal is to develop mass spectrometry (MS) instrumentation and methods for protein biophysics and structural proteomics. We wish to understand protein folding, to determine ligand binding sites, to quantify protein/ligand interactions, and to determine protein interfaces especially for systems involved in protein misfolding. A goal is a "tool box" of various methods involving covalent footprinting of proteins (e.g., H/D amide exchange, reactive radical footprinting, and specific chemical reactions). We currently emphasize H/D exchange of proteins, as monitored by ESI, as an approach to understanding protein folding and interactions, an approach that has led to a new method (PLIMSTEX) in which we use exchange and modeling to determine protein/ligand interactions. We also are investigating the reactions of OH and other radicals, generated with a short laser pulse, with proteins as a structural tool that is complementary to H/D exchange . We seek to discover new reactions for protein footprinting and to test applicability in problem solving. The instrumentation we use for this research are two Quadrupole/Time-of-Flight mass spectrometers, a 12-tesla FT-Ion Cyclotron Resonance mass spectrometer, an Orbitrap, and two 7-tesla FT ICR mass spectrometers. Analysis is by both "bottom-up" and "top-down" MS-based proteomics utilizing electrospray ionization, native spray, and MALDI and FT-ICR mass analyzers. A long-standing application area involves biochemical problems in disease (e.g., cancer, diabetes) and in immunology. Application of proteomic methods to identify antigenic peptides and to understand the basis of immune response (with E. Unanue and P. Allen) are another area of interest. We also work on methods and applications for structure determination, sequence analysis, reactivity determination, and ion-chemistry of peptides and modified pieces of oligodeoxynucleotides and DNA. We are also interested in developing Fourier transform mass spectrometry by implementing new ion-trapping methods and electrically compensated cells to afford major improvements in mass resolving power, mass measurement, and ability to store, manipulate, and react biomolecule ions and reactive species. We expect the outcome of this research to be the capability to measure accurate masses of materials at femtomole and lower levels. Our laboratory is also a NIH research resource commissioned to provide collaboration and service in mass spectrometry including molecular weight measurement, accurate mass measurement, tandem mass spectrometry experiments, and proteomics. We seek collaborations in all these areas.

Liu H, Huang RY, Chen J, Gross ML, Pakrasi HB.  Psb27, a transiently associated protein, binds to the chlorophyll binding protein CP43 in photosystem II assembly intermediates. Proc Natl Acad Sci U S A. 2011 108(45):18536-41. PMID:22031695.
 
Huang RY, Garai K, Frieden C, Gross ML.  Hydrogen/Deuterium exchange and electron-transfer dissociation mass spectrometry determine the interface and dynamics of apolipoprotein E oligomerization.Biochemistry 2011 50(43):9273-82. PMID:21899263
 
Gau B, Garai K, Frieden C, Gross ML (2011). Mass spectrometry-based protein footprinting characterizes the structures of oligomeric apolipoprotein E2, E3, and E4,Biochemistry, 50(38):8117-26. PMID: 21848287.
 
Huang RY, Rempel DL, Gross ML. HD exchange and PLIMSTEX determine the affinities and order of binding of Ca2+ with troponin C. Biochemistry 2011 50(24):5426-35. PMID:21574565
 
Zhang H, Cui W, Wen J, Blankenship RE, Gross ML.  Native electrospray and electron-capture dissociation FTICR mass spectrometry for top-down studies of protein assemblies, Anal. Chem 2011 83(14):5598-606. PMID:21612283
 
Jones LM, B Sperry J, A Carroll J, Gross ML. Fast photochemical oxidation of proteins for epitope mapping. Anal. Chem. 2011 83(20):7657-61. PMID:21894996.
 
Sperry JB, Smith CL, Caparon MG, Ellenberger T, Gross ML.  Mapping the protein-protein interface between a toxin and its cognate antitoxin from the bacterial pathogen Streptococcus pyogenes. Biochemistry 2011 50(19):4038-45. PMID:21466233.
 
Tu T, Giblin D, Gross ML.  Structural determinant of chemical reactivity and potential health effects of quinones from natural products. Chem Res Toxicol. 2011 24(9):1527-39. PMID:21721570
 
Zhang H, Shen W, Rempel D, Monsey J, Vidavsky I, Gross ML, Bose R.  Carboxyl-group footprinting maps the dimerization interface and phosphorylation-induced conformational changes of a membrane-associated tyrosine kinase. Mol Cell Proteomics. 2011 10(6):M110.005678. PMID:21142124.
 
Su D, Delaplane S, Luo M, Rempel DL, Vu B, Kelley MR, Gross ML, Georgiadis MM. Interactions of Apurinic/Apyrimidinic Endonuclease with a Redox Inhibitor: Evidence for an Alternate Conformation of the Enzyme. Biochemistry 2011 50(1):82-92 PMID:21117647