CORE--SOLID PHASE PEPTIDE SYNTHESIS AND HPLC/CZE

Salk Institute For Biological Studies, La Jolla CA

Investigators

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Abstract

This Core will synthesize and characterize peptides, and develop new HPLC- and CZE-based analytical systems to improve on resolution and efficiency. Additionally, this Core will perform circular dichroism (CD) experiments as requested. Using the Merrifield automated SPPS, Core will synthesize, each year, a total of approximately 20 peptides (10 to 50 residues in length; linear, cyclic, phosphorylated). Using the latest techniques for the preparative purification of peptide/proteins, we will provide 10 to 100 mg of purified peptides/proteins to HPLC, ion exchange chromatography on FPLC and capillary zone electrophoresis (CZE). Characterization will use mass spectroscopy (LSIMS) and Edman degradation (Core C) when necessary. These peptides will support most projects will Core C will use CZE. Peptides will be used to carry out functional and structural studies as well as to raise specific polyclonal antibodies. Finally, we intend to use some of the resources of this Core to develop new analytical techniques and explore new applications. This Core will take advantage of the P.,I.'s experience (12% effort) and that of his technical assistant as well as his fully functional laboratory for SPPS and characterization. Laboratory space equipped with hoods and other small equipment is available. Major instruments available include an LSI Mass Spectrometer Model HX-110 and a 60 MHz NMR from JEOL, a Beckman P/ACE model 2050 CZE, three peptide synthesizers from Beckman, analytical and preparative HPLCs and lyophilizers. The P.I. will chair the users' committee responsible for setting priorities. It is expected that Core D will be able to operate on a first come, first served basis. In cases where priority will have to be determined, the decision reached by the majority of the users' committee (excluding the chair) will be implemented. Synthesis and purification of peptides will be carried out by M. Odrowaz-Sypniewski (30% effort). Quality control and method development using HPLC, FPLC and CZE will be carried out by D. Kirby (15% effort). MS analyses (15% effort) will be carried out by Dr. A. Craig (15% effort). With the consent of the concerned P.I.s, excess peptides will be distributed free of charge to any NIH sponsored researcher as we have done in the past. Recently, there has been a significant need for circular dichroism spectroscopy. We have been responsible for the instrument for more than ten years and will provide analyses to investigators on this Program Project.