Funds are requested for purchase of equipment to support Difference Gel Electrophoresis (DIGE) (scanner, software and supporting items). While Columbia University has resources for identification of proteins it has no capability in comparative proteomics. The proposed integrated combination of hardware and software will allow the university to leverage existing capabilities by adding a workflow to evaluate relative differential expression of proteins. A group of 35 major users listed in this proposal include NIH grantees drawn from the main (Morningside Heights), the uptown (Medical Center) campuses of Columbia as well as from other universities. Response from other campuses indicates that this lab is on its way to becoming a regional center. Other users will undoubtedly be continuously added to this list as full participants in future years. The Department of Biological Sciences on the main campus recruited the Principal Investigator in September 2006 to manage this new collaborative DIGE laboratory. The Principal Investigator has five years of hands-on experience with DIGE, and for this proposal has generated preliminary data for 9 of the 35 collaborating laboratories. Data from preliminary experiments are presented and demonstrate that the technique is appropriate and effective for detecting differential expression of proteins for these NIH-funded projects. Five of these research groups have performed (in some cases extensive) corroborative experiments in their laboratories validating the DIGE results using non-proteomic methods. Publications representing some of these results are in press or submitted to high impact journals. Other alternative technologies have merits and can detect complementary sets of differentially expressed proteins, but these do not as easily leverage existing resources, and are not appropriate to many of the studies that we are planning. Studies have shown that the skills and experience of the laboratory with the particular technique used is more important than the technique itself. The department and university are supporting the establishment of the DIGE laboratory through long-term salary support, startup funds for supplies, salaries and robotics equipment to support DIGE, cost-share funds for a new mass spectrometer, funds for laboratory renovations, funds for continuing service contracts and space allocated to house the requested equipment and supporting activities. The laboratory also enjoys significant infrastructural support from the department, and support from an advisory committee to help manage the equipment.
This equipment will support research in a wide range of topics in basic research and in the understanding of disease processes. Proteins will be compared among different physiological states or disease processes.
| Brown, Lewis M (2014) Quantitative shotgun proteomics with data-independent acquisition and traveling wave ion mobility spectrometry: a versatile tool in the life sciences. Adv Exp Med Biol 806:79-91 |
| Higashi, Dustin L; Biais, Nicolas; Weyand, Nathan J et al. (2011) N. elongata produces type IV pili that mediate interspecies gene transfer with N. gonorrhoeae. PLoS One 6:e21373 |
| Oswald, Elizabeth S; Brown, Lewis M; Bulinski, J Chloe et al. (2011) Label-free protein profiling of adipose-derived human stem cells under hyperosmotic treatment. J Proteome Res 10:3050-9 |
| Poyurovsky, Masha V; Katz, Chen; Laptenko, Oleg et al. (2010) The C terminus of p53 binds the N-terminal domain of MDM2. Nat Struct Mol Biol 17:982-9 |
