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The tryptophan fluorescence of Tetracarbidium conophorum agglutinin II and a solution-based assay for the binding of a biantennary glycopeptide
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The tryptophan fluorescence of Tetracarbidium conophorum agglutinin II and a solution-based assay for the binding of a biantennary glycopeptide

Martha P. Brown, Dimitri Toptygin, K.B. Lee, Theresa Animashaun, R.C. Hughes, Y.C. Lee and Ludwig Brand
Protein Journal, Vol.17(2), pp.149-159
1998

Abstract

Fluorescence Glycopeptide Lectin Lifetime Tryptophan Analytical Chemistry Bioengineering Biochemistry Organic Chemistry
The plant lectin Tetracarbidium conophorum agglutinin II binds to glycoproteins and glycopeptides in a structurally specific manner [Animashaun et al., (1994) Glycoconjugate J. 11, 299-303]. We have characterized the steady-state and time-resolved fluorescence of the tryptophan residues of this lectin. The fluorescence (λ ex = 295 nm, λ em = 350 nm) decay is complex and can be described by four decay times with the following values: τ 1 = 7.4 nsec, α 1 = 0.22; τ 2 = 2.9 nsec, α 2 = 0.25; τ 3 = 1.0 nsec, α 3 = 0.34; τ 4 = 0.2 nsec, α 4 = 0.18. The addition of a biantennary glycopeptide (Chemical Equation Presented) to the lectin results in a quench and an 8 nm blue shift of the emission spectrum. The effect is saturable, and is described by an association constant of 1.8 x 10 5 M -1 . The tryptophan fluorescence of Tetracarbidium conophorum agglutinin II may therefore be utilized to characterize thermodynamically the binding interactions between this lectin and complex glycoprotein. © 1998 Plenum Publishing Corporation.

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