Bio-Organic Biomedical Mass Spectrometry Resource

Bio-Organic Biomedical Mass Spectrometry Resource

Research Emphasis

The resource focuses on mass spectrometric techniques for sequencing and quantitating peptides and structural characterization of modified proteins and glycoconjugates; microsample handling and mass spectrometric analysis at the femtomole to attomole level, such as in the identification of proteins separated by two-dimensional (2-D) gel electrophoresis using silver and fluorescence detection; cotranslational and posttranslational modifications of proteins, including glycosylation, phosphorylation, sulfation, and acylation; and xenobiotic modification, such as that arising from mechanism-based enzyme inhibition and drug-protein adduct formation. Methodology includes matrix-assisted laser desorption ionization with high-energy collision-induced dissociation analysis and capillary high-performance liquid chromatography (HPLC) nanoflow rate electrospray tandem mass spectrometry.

Current Research

Molecular and cellular proteomics, drug metabolic activation, and protein adduct identification; studies of protein machines, complexes and signaling cascades, and structural biology; studies of Conus peptides that are blockers of ion channels and neurotransmission. In addition, development of multidimensional ion exchange and reverse-phase separations of affinity-separated, isotopically labeled peptides from cell culture and in vivo systems using new acid-cleavable, carbon-13-coded ICAT reagents. Development of complementary offline chromatographic [matrix-assisted laser desorption ionization (MALDI-(CIDMS)] and online (ESI-CIDMS) strategies for comprehensive detection and sequencing of ICAT-labeled systems. Development of electrospray ionization and MALDI and tandem technologies for determination of biomolecular and structural studies of proteins, noncovalent and covalent protein assemblages, heterobiopolymers, and glycoconjugates.

Resource Capabilities

Instruments

PerSeptive Biosystems MALDI Voyager Elite and Voyager Elite STR DE reflectron time-of-flight (TOF) mass spectrometers, Mariner ES oaTOF mass spectrometers, Sciex API 300 ES triple quadrupole mass spectrometer, nanoflow rate electrospray quadrupole orthogonal acceleration TOF mass spectrometer (QqTOFMS)-PE Sciex PULSAR/Q STAR, automated ABI MALDI TOF TOFMS(ABI 4700 Proteomics Analyzer) with 24-plate cassette sample system, and VG 70S mass spectrometer.

Special Features

MALDI high-energy CID capability for de novo protein sequence determination and structural analysis. Online capillary HPLC electrospray mass spectrometry and offline capillary HPLC matrix-assisted laser desorption tandem mass spectrometry for identification of one-dimensional and 2-D gel spots as well as solution protein mixtures from immunoprecipitates, fusion baits, and tandem affinity tags, etc. Bioinformatics tools such as MS-Fit, MS-Tag, and MS-Homology. Web-accessible algorithms for gene and EST database interrogation with mass spectral data via ProteinProspector.