We propose to develop a resource center with a multidisciplinary, integrated approach towards protein structural analysis, incorporating new, high speed, high sensitivity analytical methods with sample handling techniques. The goals include the development of tandem microcolumn (greater than 1mm ID) LC systems for purification, new generations of more sensitive Edman microsequencers, carboxyl terminal sequencers with novel membranes for sample immobilization, and new methods for sample introduction into mass spectrometers with a variety of sample ionization sources. The initial goal will be to perform routine protein structure analysis at the 1 picomole level an then at the 10-100 fmole level. Integrated sample handling/instrumentation will include protein purification, peptide mapping, and structural analysis using microcolumn LC, SDS-gel electrophoresis, and mass spectrometry. Mass spectrometers include laser desorption/time of flight, electrospray and fast atom bombardment (FAB)/triple quadrupole, and FAB/magnetic sector instruments, providing capabilities for high mass, and MS/MS measurements under a variety of conditions with high sensitivity and speed. Our existing facility is equipped with state-of-the-art instrumentation for protein isolation/structural analysis, has a proven record of accomplishments and innovation in these areas, and is staffed with a group of highly interactive, experienced, interdisciplinary scientists. In short, we have an ideal setting for further method/instrument development for the creation of a unique protein resource center.
