Accession Number : ADA183190

Title :   Design and Synthesis of Semisynthetic Enzymes.

Descriptive Note : Annual rept. Aug 86-Jul 87,

Corporate Author : CALIFORNIA UNIV BERKELEY DEPT OF CHEMISTRY

Personal Author(s) : Schultz,Peter G

PDF Url : ADA183190

Report Date : 28 Jul 1987

Pagination or Media Count : 22

Abstract : We propose to develop methodology which will for the first time enable specific amino acid residues in proteins to be site-specifically replaced with synthetic amino acid analogues. Amino acids with novel steric, electronic, or spectroscopic properties will then be used to probe structure-function relationships in proteins which lead to receptor/substrate recognition and/or catalysis. We are pursuing a combined chemical/genetic strategy toward this objective. Our approach requires replacement of the codon for the target amino acid with an amber nonsense codon (5'-TAG-3) via oligonucleotide-directed mutagenesis. A nonsense suppressor RNA that recognizes this unique blank codon is being constructed by both anticodon loop replacement and by chemical synthesis and overproduction (E. coli) of an amber suppressor gene, using known chemical/biological procedures. This suppressor RNA will then be chemically aminoacylated with novel amino acids by existing chemical methods, thereby avoiding the natural specificity of the aminoacyl RNA synthetases. The aminoacylated RNA will be used as a delivery vehicle in vitro for placing novel synthetic amino acids site-specifically into proteins. The methodology described above will be developed and optimized with the enzyme, Beta-lactamase, which catalyzes the hydrolysis of the Beta-lactam antibiotics penicillin and cephalosporin.

Descriptors :   *GENETICS, *SYNTHESIS(CHEMISTRY), *ENZYMES, *AMINO ACIDS, GENES, SUPPRESSORS, CATALYSIS, CHEMICALS, STRATEGY, BIOLOGY, DELIVERY, IN VITRO ANALYSIS, ANTIBIOTICS, PENICILLINS, MUTATIONS, NUCLEOTIDES, HYDROLYSIS, PROTEINS, RESIDUES, RIBONUCLEIC ACIDS, LOOPS, REPLACEMENT, RECOGNITION, SENSE ORGANS, SUBSTRATES, SPECTROSCOPY, SYNTHESIS, TARGETS

Subject Categories : Biochemistry
      Genetic Engineering and Molecular Biology

Distribution Statement : APPROVED FOR PUBLIC RELEASE