[58] Determination of the tryptophan content of proteins with N-bromosuccinimide

Spande, Thomas F.; Witkop, Bernhard

doi:10.1016/s0076-6879(67)11060-4

Cited by 489 publications

(248 citation statements)

References 14 publications

Supporting

Mentioning

240

Contrasting

Order By: Relevance

“…Increasing amounts of Nbromosuccinimide led to a decrease in the absorbance at 280nm but an increase at 250 nm, showing that tryptophan residues were transformed to their oxidole tives. 19 ) of the purified acidic lectin was specifically inhibited by D-galactose and related sugars. Among the tested sugars, N-acetyl-D-galactosamine was the best inhibitor, causing 100% inhibition of the agglutination at a concentration of 3.1 mM.…”

Section: Inhibition Of Agglutination By Sugarsmentioning

confidence: 95%

Purification and Characterization of the Acidic Lectin from Winged Bean Seeds

Higuchi¹,

Ohtani²,

Iwai³

1986

Agricultural and Biological Chemistry

View full text Add to dashboard Cite

Section: Inhibition Of Agglutination By Sugarsmentioning

confidence: 95%

Purification and Characterization of the Acidic Lectin from Winged Bean Seeds

Higuchi¹,

Ohtani²,

Iwai³

1986

Agricultural and Biological Chemistry

View full text Add to dashboard Cite

“…2) and there were no further changes in the absorbance of the sample at 280 nm. The number of tryptophans oxidized by NBS was calculated by the absorbance method (Spande & Witkop, 1967).…”

Section: Chemical Modi9cation Of Cbdcpxmentioning

confidence: 99%

Probing the role of tryptophan residues in a cellulose‐binding domain by chemical modification

et al. 1996

View full text Add to dashboard Cite

The cellulose-binding domain (CBD,,,) of the mixed function glucanase-xylanase Cex from Cellulomonas fimi contains five tryptophans, two of which are located within the p-barrel structure and three exposed on the surface (Xu GY et al., 1995, Biochemistry 346993-7009). Although all five tryptophans can be oxidized by N-bromosuccinimide (NBS), stopped-flow measurements show that three tryptophans react faster than the other two. NMR analysis during the titration of CBDcex with NBS shows that the tryptophans on the surface of the protein are fully oxidized before there is significant reaction with the two buried tryptophans. Additionally, modification of the exposed tryptophans does not affect the conformation of the backbone of CBDcex, whereas complete oxidation of all five tryptophans denatures the polypeptide. The modification of the equivalent of one and two tryptophans by NBS reduces binding of CBDcex to cellulose by 70% and 90%, respectively. This confirms the direct role of the exposed aromatic residues in the binding of CBDcex to cellulose. Although adsorption to cellulose does afford some protection against NBS, as evidenced by the increased quantity of NBS required to oxidize all of the tryptophan residues, the polypeptide can still be oxidized completely when adsorbed. This suggests that, whereas the binding appears to be irreversible overall [Ong E et al., 1989, Bio/Technology 76044071, each of the exposed tryptophans interacts reversibly with cellulose.

show abstract

“…METHODS 3.1. Chemical modification with Nbromosuccinimide The modification of G2 in the presence of specific ligands was performed essentially as previously described (9,27). Aliquots (200-400 lal) of 10 mM-NBS were sequentially added to a Millipore-filtered solution (50 ml) of G2 (15 ~tM) containing either acarbose (0.15 mM) or maltose (56 mM) in 50 mM-sodium acetate pH 4.3.…”

Section: Introductionmentioning

confidence: 99%

Identification of an essential tryptophanyl residue in the primary structure of glucoamylase G2 from aspergillus niger

Clarke

Svensson

1984

Carlsberg Res. Commun.

View full text Add to dashboard Cite

Enzymatically active, N-bromosuccinimide oxidized glucoamylase G2 (EC 3.2.1.3) was prepared in the presence of the inhibitor acarbose and inactive, oxidized G2 with capacity to bind substrate was prepared in the presence of maltose. Four out of 15 tryptophanyl residues were oxidized in the first derivative and five in the latter. In order to identify this fifth and essential residue, tryptie fragments of the two G2 derivatives were isolated. The peptide fragment from the inactive G2 der/vative containing the additional oxindolealaniae residue was Tocated in HPLC chromatograms by UV-absorption measurements. Normal and second derivative UV-spectra, amino acid composition and NH_,-terminal sequence of the isolated fragment led to identification of Trp(120) as a residue essential for activity. A short homologous amino acid sequence precedes both this Trp(120) and the Trp(83) which is involved in substrate binding in Taka-amylase A (J. Biochem. 95, 697-702 (1984)).

show abstract

[58] Determination of the tryptophan content of proteins with N-bromosuccinimide

Cited by 489 publications

References 14 publications

Purification and Characterization of the Acidic Lectin from Winged Bean Seeds

Purification and Characterization of the Acidic Lectin from Winged Bean Seeds

Probing the role of tryptophan residues in a cellulose‐binding domain by chemical modification

Identification of an essential tryptophanyl residue in the primary structure of glucoamylase G2 from aspergillus niger

Contact Info

Product

Resources

About