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Spectrofluorometric Properties of N-Terminal Domain of Lumazine Protein from Photobacterium leiognathi

  • Kang, Kyoung-Suk (Department of Biochemistry, Chungnam National University) ;
  • Kim, So-Young (Department of Biochemistry, Chungnam National University) ;
  • Lee, Jung-Hwan (Department of Biochemistry, Chungnam National University) ;
  • Nam, Ki-Seok (Department of Biochemistry, Chungnam National University) ;
  • Lee, Eui Ho (Department of Biochemistry, Chungnam National University) ;
  • Lee, Chan Yong (Department of Biochemistry, Chungnam National University)
  • Received : 2013.01.16
  • Accepted : 2013.03.13
  • Published : 2013.06.20
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Abstract

Lumazine protein is a member of the riboflavin synthase superfamily and the intense fluorescence is caused by non-covalently bound to 6,7-dimethyl 8-ribityllumazine. To figure out the binding modes and the structure of the N-terminal domain of lumazine protein, the wild type of protein extending to amino acid 118 (N-LumP 118 Wt) and mutants of N-LumP 118 V41W, S48W, T50W, D64W, and A66W from Photobacterium leiognathi were purified. The biochemical properties of the wild type and mutants of N-LumP 118 proteins were analyzed by absorbance and fluorescence spectroscope. The peak of absorbance and fluorescence of lumazine ligand were shifted to longer wavelength on binding to N-LumPs. The observed absorbance value at 410 nm of lumazine bound to N-LumP 118 proteins indicate that one mole of N-LumP 118 proteins bind to one mole of ligand of lumazine. Fluorescence analysis show that the maximum peak of fluorescence of N-LumP S48W was shifted to the longest wavelength by binding with 6,7-dimethyl 8-ribityllumazine and was shown to the greatest quench effect by acrylamide among all tryptophan mutants.

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