caa a22 4500 39755706X CHVBK 20180308164805.0 cr unu---uuuuu 161202e199607 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021399 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021399 Purification and Characterization of the Proteus vulgaris BlaA Protein, the Activator of the β-Lactamase Gene [Elektronische Daten] [Koji Ishiguro, Kazunori Sugimoto] Induction of the expression of the β-lactamase gene, blaB, is regulated by the blaA gene located just upstream of blaB in the opposite direction in Proteus vulgaris. The expression of the blaA gene is negatively autoregulated by its own product BlaA, the activator of the blaB gene. The P. vulgaris BlaA protein shares high amino acid homology with the LysR family members, which are prokaryotic transcriptional activators that possess a putative helix-turn-helix DNA binding motif. To characterize its function, we purified the BlaA protein to homogeneity from Escherichia coli carrying the expression plasmid of the blaA gene driven by the tac promoter. The gel shift assay and DNasel footprinting showed that purified BlaA specifically bound to the blaA promoter region, which resides immediately upstream of that of blaB. The binding region contained an inverted repeat, including a T-N11-T sequence which is similar to the LysR motif (T-N11-A) that is conserved in some LysR family members [Goethals et al. (1992) Proc. Natl. Acad. Sci USA 89, 1646-1650]. We also showed that the BlaA protein forms a dimer in solution, using glycerol gradient centrifugation and glutaraldehyde crosslinking. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence β-lactamase nationallicence BlaA nationallicence DNA binding protein nationallicence LysR family nationallicence Ishiguro Koji Division of Chemistry, Graduate School of Science, Hokkaido University, Kita-ku, Sapporo 060 aut Sugimoto Kazunori Division of Chemistry, Graduate School of Science, Hokkaido University, Kita-ku, Sapporo 060 aut The Journal of Biochemistry Oxford University Press 120/1(1996-07), 98-103 0021-924X 120:1<98 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021399 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021399 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ishiguro Koji Division of Chemistry, Graduate School of Science, Hokkaido University, Kita-ku, Sapporo 060 aut NATIONALLICENCE 700 1- Sugimoto Kazunori Division of Chemistry, Graduate School of Science, Hokkaido University, Kita-ku, Sapporo 060 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/1(1996-07), 98-103 0021-924X 120:1<98 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford