title: DNA-binding specificity of the Lon protease alpha-domain from Brevibacillus thermoruber WR-249.
abstract: Lon protease has been well studied in many aspects; however, the DNA-binding specificity of Lon in prokaryotes has not been clearly identified. Here we examined the DNA-binding activity of Lon protease alpha-domains from Brevibacillus thermoruber (Bt), Bacillus subtilis (Bs), and Escherichia coli (Ec). MALDI-TOF mass spectroscopy showed that the alpha-domain from Bt-Lon binds to the duplex nucleotide sequence 5'-CTGTTAGCGGGC-3' (ms1) and protected it from DNase I digestion. Surface plasmon resonance showed that the Bt-Lon alpha-domain binds with ms1 double-stranded DNA tighter than Bs- and Ec-Lon alpha-domains, whereas the Bt-Lon alpha-domain has dramatically lower affinity for double-stranded DNA with 0 and 50% identity to the ms1 binding sequence. Our results indicated that Bt-Lon alpha-domain plays a critical role with ms1 sequence in the DNA-binding specificity.
authors: Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan.
authors: Yu-Ching
authors: YC
authors: Lin
authors: Huai-Cheng
authors: HC
authors: Lee
authors: Iren
authors: I
authors: Wang
authors: Chun-Hua
authors: CH
authors: Hsu
authors: Jiahn-Haur
authors: JH
authors: Liao
authors: Alan Yueh-Luen
authors: AY
authors: Lee
authors: Chinpan
authors: C
authors: Chen
authors: Shih-Hsiung
authors: SH
authors: Wu
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology*; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000201:enzymology; D008969:Molecular Sequence Data; D049070:Protease La / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D004268:DNA-Binding Proteins; D004247:DNA; D049070:Protease La