The Protein Structure facility, located in the Department of Cell Biology and Anatomy, assists with the purification of proteins, peptide mapping from in situ digestions in gels or from PVDF membranes, microsequencing and MALDI-TOF mass spectroscopic analysis. Thisresource has an established track record in primary gene discovery, proteomics and characterization of post-translational modification.
By applying techniques that allow structural analysis of proteins in the relatively low pmol range, this facility has played a major role in identifying new genes. The cloning of the platelet derived growth factor receptor, basic FGF receptor and key components of the neurosecretory pathway have been accomplished by developing protein purification, fragmentation and sequencing procedures appropriate for small amounts of material within the 5 -to 10-pmol range.
The advent of complete genomic databases for many organisms have opened the way to a new strategy for defining cellular states that establish both normal and pathological activities. Protein chemistry for gene discovery is being replaced by protein analytical and sequencing techniques. Coupled with a genomic database (proteomics), these have become a powerful tool in studying gene expression. With most of these new approaches, only small amounts of sequence are needed to identify a protein detected by high resolution 1-D and 2-D gene separation techniques. Analysis of proteolytic digests of a protein by mass spectroscopy (MS), and effective algorithms for screening the products predicted from the genomic database, provide sensitive and generally accurate identification of the protein. Using these proteomic strategies, the Protein Structure facility has identified the protein structure of the regulatory particle of the yeast proteosome, the subassembly of this structure and completed MS analysis of protein subunits of a mutated form of the yeast 26s proteosome.
To address the extensive post-translational modification that primary gene products undergo, the Protein Structure facility has developed approaches for determining sites of glycosylation, proteolytic processing and phosphorylation of a natural protein at the low pmol level. The facility’s proven ability to identify gene products and define post-translational modifications enable it to deliver an integrated approach for analyzing gene expression and regulatory pathways.
Victor A. Fried, Ph.D.
Professor of Cell Biology and Anatomy
New York Medical College
Department of Cell Biology and Anatomy
Basic Sciences Building
Valhalla, NY 10595