The acquisition of protein sequencing instrumentation is sought for the Genetic Engineering Facility of the Biotechnology Center at the University of Illinois. The research requiring this instrumentation is very diverse as reflected in the range of research projects that are described. Several investigators require the protein sequencer for problems involving the structure of enzyme active sites. The residue modified by site-specific reagents acting on the enzyme can be identified. Analysis of the amino acid sequence of monoclonal antibodies with differing affinities for a single hapten is required to define the components of the antigen binding site which determine the binding affinity. When combined with X-Ray crystallographic data on these antibodies, these studies will serve as the basis for future molecular modeling and genetic engineering programs to construct antibodies of defined specificity. The amino acid changes in active sites which result from site- directed mutagenesis of DNA must be confirmed by sequencing. Knowledge of N-terminal amino acid sequence of proteins and peptides are required to identify open reading frames of sequenced DNA. Often very small amounts of protein are available for these types of experiments and state-of-the-art instrumentation with sensitivity in the range of 10 pmoles or less is required. A unique application of this protein sequencer is in studies on the specificity of RNA translation in eucaryotic cells. Presence of this equipment on campus in a centralized facility will enable faculty to carry out many experiments that otherwise would be too expensive or beyond the capabilities of laboratory personnel. It also provides the most effective use of available equipment and avoids duplication of costly instrumentation in individual laboratories. The Biotechnology Center and the Graduate College will provide the laboratory space and full support facilities for operation of these instruments. On going maintenance and the presence of trained technical personnel will be assured by the Graduate College.
