PROTEIN-PEPTIDE SEQUENCING FACILITY

点击翻译按钮获取中文摘要

The objective of this proposal is to establish a "state of the art facility" for the analysis and characterization of proteins and peptides using automated microsequencing techniques, determination of amino acid composition, high performance liquid chromatography and DNA oligonucleotide synthesis. Our goal Is to establish a centralized protein structure facility housed in one laboratory. The major instrumentation requested is: (i) an Applied Biosystems Pulsed Liquid Phase Protein/Peptide Sequencer with on-line identification of phenylthlohydantoin derivatives; (ii) an Applied Biosystems Derivatizer Amino Acid Analyzer System for the determination of amino acid composition; iii) a Waters High Performance Liquid Chromatography System for preparative and analytical purification of proteins, peptides, phenylthiohydantoins and oligonucleotides and (iv) an Applied Biosystems DNA synthesizer for the preparation of oligonucleotides. This instrumentation and a recently acquired peptide synthesizer will provide investigators with three types of essential methodologies: protein sequencing, peptide synthesis and DNA synthesis. The facility will ensure that investigators can start at the protein, messenger RNA or gene levels to generate "genes from proteins" or "proteins from genes". The availability of this protein structure instrumentation is essential for continued productivity and optimal progress in the broad spectrum of research programs at Albert Einstein College of Medicine that are addressing fundamental problems in biological regulation. Presently, this Institution possesses one eight year old Beckman 890 C spinning-cup sequencer; although this instrument is still operational it is not sufficiently sensitive or reliable for modern sequencing applications. This application (a) documents the need for an up-dated centralized facility which will be available to our entire scientific community; (b) describes the feasibility of installing, operating and maintaining such a facility and (c) outlines a series of projects that require protein sequencing and which will make immediate use of the facility.