Core B
MASS SPECTROMETRY
Catherine Costello, PhD |
Core Leader: Catherine E. Costello, Ph.D., Research Professor of Biochemistry and Biophysics
Mass Spectrometry Core B provides support for all projects with state-of-the-art mass spectrometry (MS) and atomic force microscopy (AFM). MS Core B will work closely with the Protein Purification Core A.
The BUSM Center for Biological Mass Spectrometry comprises the NIH-NCRR Research Resource for Mass Spectrometry in Biology and Medicine, the Cardiovascular Proteomics Center and the Shared Instrumentation Laboratory. The Center has available electrospray ionization (ESI) on triple quadrupole, quadrupole orthogonal time-of-flight (Q-o-TOF), and ion cyclotron resonance Fourier transform (FTICR) mass spectrometers, and ultraviolet and infrared matrix-assisted laser desorption/ionization (MALDI) on Q-o-TOF, reflectron TOF and FTICR mass spectrometers, and an atomic force microscope, as well as a highly trained staff and a commitment to the development and application of methodology for biopolymer structural analysis.
Samples include protein (and other biopolymer) fractions separated by prior or on-line chromatography, as well as amyloid fibrils, laser capture microdissection samples and spots from SDS/PAGE and IEF gels that are subjected to in-gel digestion prior to MS and multistage MS analysis. Results are compared to protein sequences predicted from gene sequences and with on-line databases and/or interpreted manually to yield complete primary sequences whether or not genetic information is available.
Core B supports Project 1 with molecular weight determinations and sequence verifications of expressed proteins, assessment of purity and elucidation of variations in amino acid sequence and posttranslational modifications, including structural determinations of N- and O-linked glycans. For Project 2, the same types of information are provided for clinically-derived proteins and oligosaccharides. Peptides and proteins from Project 3 are identified by MS and MS/MS, if required to supplement data from Core A. In collaboration with Projects 1 and 2, fibril deposits and fibril growth are being monitored by AFM.
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