Sugar binding effects on the enzymatic reaction and conformation near the active site of pokeweed antiviral protein revealed by fluorescence spectroscopy

Hiromichi Nakashima, Yukihiro Fukunaga, Ryosuke Ueno, Etsuko Nishimoto

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

In various trials for elucidating the physiological function of pokeweed antiviral protein (PAP), studies on the interaction with sugar are essential. The fluorescence titration curves showed that PAP retained the strong affinity against N-acetylglucosamine (NAG) and two sites in one PAP molecule co-operatively participated in the binding. In the complex of PAP with NAG, Trp208 located at the entrance lid site of substrate came closer to Tyr72 about 0.3 Å. Furthermore, the fluorescence anisotropy decay measurement demonstrated that the segmental rotation of Trp208 was enlarged by the binding of PAP with NAG. Such conformational changes around the active site closely correlate with the enzymatic activity of PAP. The N-glycosidase activity of PAP was enhanced more than two times in the presence of NAG. The obtained results consistently suggested the enzymatic activity of PAP would be regulated through the conformation change near the active site induced by the binding with NAG.

Original languageEnglish
Pages (from-to)951-958
Number of pages8
JournalJournal of Fluorescence
Volume24
Issue number3
DOIs
Publication statusPublished - Jan 1 2014

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Fluorescence Spectrometry
Fluorescence spectroscopy
Sugars
Conformations
Catalytic Domain
Acetylglucosamine
Fluorescence
interaction
Fluorescence Polarization
pokeweed antiviral protein
Glycoside Hydrolases
Titration
Anisotropy
Molecules
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Spectroscopy
  • Clinical Biochemistry

Cite this

Sugar binding effects on the enzymatic reaction and conformation near the active site of pokeweed antiviral protein revealed by fluorescence spectroscopy. / Nakashima, Hiromichi; Fukunaga, Yukihiro; Ueno, Ryosuke; Nishimoto, Etsuko.

In: Journal of Fluorescence, Vol. 24, No. 3, 01.01.2014, p. 951-958.

Research output: Contribution to journalArticle

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