Protein Structure Analysis and Synthesis Facility

Dunn, Ben

Abstract

This proposal requests funding to update our Protein Sequence Facility through the acquisition of a new amino acid analyzer and through a microsequencing update on our """"""""Spinning Cup"""""""" Sequencer. Our current efforts are handicapped by our lack of a fast, sensitive analyzer capable of providing accurate data on small amounts of material. Many of our colleagues are now studying the biological response of very small amounts of proteins obtained by electrophoresis or other techniques. It is essential that we be able to provide reliable analytical data on these samples. The microsequencing update will enable us to significantly improve our operation by providing rapid conversion of ATZ- to PTH-amino acids. It will also provide improvements in operation to enable us to reach the 200 picomole level in our sequencing operation. We are also requesting funds for the purchases of a new oligopeptide synthesizer and a new oligonucleotide synthesizer. Each of these units will make a significant contribution to our overall program by allowing us to prepare new fragments for biochemical study. Oligopeptides are being used as synthetic antigens as well as enzyme substrates. In other cases, the biological activity of these fragments is being measured directly. Oligonucleotides are useful in many projects as probes for complementary regions of DNA. By synthesis of specific oligonucleotides designed from the primary amino acid sequence of a protein of interest, the oligonucleotide probe approach will permit the detection and isolation of specific genes or gene fragments.