Abstract

Structure-function studies are to be conducted with anthranilate synthase from Serratia marcescens. Group-specific reagents will be used to inactivate the enzyme and identify residues essential for NH3-dependent activity. Large amounts of E. coli amidophosphoribosyl transferase will be purified from a strain carrying a purF recombinant plasmid. The essential active site cystein will be labeled with 6-diazo-5-oxonorleucine, and an active site tryptic peptide will be isolated and sequenced. The nucleotide sequence of the purF gene will be determined, thus allowing the glutamine active site region to be placed in the sequence and a comparison with other glutamine amidotransferases to be made.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM024658-15
Application #
3272424
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1977-07-01
Project End
1987-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
15
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Purdue University
Department
Type
Earth Sciences/Resources
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907

  Publications

Sinha, Sangita C; Krahn, Joseph; Shin, Byung Sik et al. (2003) The purine repressor of Bacillus subtilis: a novel combination of domains adapted for transcription regulation. J Bacteriol 185:4087-98
Bera, Aloke Kumar; Zhu, Jianghai; Zalkin, Howard et al. (2003) Functional dissection of the Bacillus subtilis pur operator site. J Bacteriol 185:4099-109
Bera, A K; Smith, J L; Zalkin, H (2000) Dual role for the glutamine phosphoribosylpyrophosphate amidotransferase ammonia channel. Interdomain signaling and intermediate channeling. J Biol Chem 275:7975-9
Bera, A K; Chen, S; Smith, J L et al. (2000) Temperature-dependent function of the glutamine phosphoribosylpyrophosphate amidotransferase ammonia channel and coupling with glycinamide ribonucleotide synthetase in a hyperthermophile. J Bacteriol 182:3734-9
Weng, M; Zalkin, H (2000) Mutations in the Bacillus subtilis purine repressor that perturb PRPP effector function in vitro and in vivo. Curr Microbiol 41:56-9
Bera, A K; Chen, S; Smith, J L et al. (1999) Interdomain signaling in glutamine phosphoribosylpyrophosphate amidotransferase. J Biol Chem 274:36498-504
Li, S; Smith, J L; Zalkin, H (1999) Mutational analysis of Bacillus subtilis glutamine phosphoribosylpyrophosphate amidotransferase propeptide processing. J Bacteriol 181:1403-8
Rappu, P; Shin, B S; Zalkin, H et al. (1999) A role for a highly conserved protein of unknown function in regulation of Bacillus subtilis purA by the purine repressor. J Bacteriol 181:3810-5
Sinha, S; Rappu, P; Lange, S C et al. (1999) Crystal structure of Bacillus subtilis YabJ, a purine regulatory protein and member of the highly conserved YjgF family. Proc Natl Acad Sci U S A 96:13074-9
Lu, F; Schumacher, M A; Arvidson, D N et al. (1998) Structure-based redesign of corepressor specificity of the Escherichia coli purine repressor by substitution of residue 190. Biochemistry 37:971-82

Showing the most recent 10 out of 79 publications