Formation of δ1-Acetoxytryptophan-62 in the Oxidation of Tryptophan-62 of Hen Egg-White Lysozyme by N-Bromosuccinimide in Acetate Buffer

Raymond S. Norton, Adam Allerhand

Research output: Contribution to journalArticleResearchpeer-review

23 Citations (Scopus)

Abstract

The reaction of equimolar amounts of N-bromosuccinimide and hen egg-white lysozyme in acetate buffer, under the conditions of Hayashi et al. (Hayashi, K., Imoto, T., Funatsu, G. and Funatsu, M. (1965), J. Biochem. (Tokyo) 58, 227), yields a protein mixture that has a time-dependent 13C-NMR spectrum. The initial natural-abundance 13C-NMR spectrum indicates the presence of about equal amounts of [oxindolealanine-62]lysozyme and [δ1-acetoxytryptophan-62] lysozyme. The latter converts to [oxindolealanine-62] lysozyme w ith a half-life of about 2 days at 25 °C and pH 3.9. Two observations indicate that the source of the acetylgroup of δ1-acetoxytryptophan-62 is the acetate buffer. First, the spectrum of a lysozyme sample treated with N-bromosuccinimide in the presence of [1-13C]acetate yields a verystrong acetyl ester carbonyl resonance. The time dependence of the intensity of this resonance yields a half-life of 44 h for [δ1-acetoxytryptophan-62]lysozyme. Second, the initial natural-abundance 13C-NMR spectrum of a lysozyme sample treated with A′-bromosuccinimide in the absence of acetate indicates essentially complete conversion of tryptophan-62 into oxindolealanine.

Original languageEnglish
Pages (from-to)3438-3445
Number of pages8
JournalBiochemistry
Volume15
Issue number16
DOIs
Publication statusPublished - 1 Aug 1976
Externally publishedYes

Cite this

@article{5e87da19adf94a569a4da0223365f45d,
title = "Formation of δ1-Acetoxytryptophan-62 in the Oxidation of Tryptophan-62 of Hen Egg-White Lysozyme by N-Bromosuccinimide in Acetate Buffer",
abstract = "The reaction of equimolar amounts of N-bromosuccinimide and hen egg-white lysozyme in acetate buffer, under the conditions of Hayashi et al. (Hayashi, K., Imoto, T., Funatsu, G. and Funatsu, M. (1965), J. Biochem. (Tokyo) 58, 227), yields a protein mixture that has a time-dependent 13C-NMR spectrum. The initial natural-abundance 13C-NMR spectrum indicates the presence of about equal amounts of [oxindolealanine-62]lysozyme and [δ1-acetoxytryptophan-62] lysozyme. The latter converts to [oxindolealanine-62] lysozyme w ith a half-life of about 2 days at 25 °C and pH 3.9. Two observations indicate that the source of the acetylgroup of δ1-acetoxytryptophan-62 is the acetate buffer. First, the spectrum of a lysozyme sample treated with N-bromosuccinimide in the presence of [1-13C]acetate yields a verystrong acetyl ester carbonyl resonance. The time dependence of the intensity of this resonance yields a half-life of 44 h for [δ1-acetoxytryptophan-62]lysozyme. Second, the initial natural-abundance 13C-NMR spectrum of a lysozyme sample treated with A′-bromosuccinimide in the absence of acetate indicates essentially complete conversion of tryptophan-62 into oxindolealanine.",
author = "Norton, {Raymond S.} and Adam Allerhand",
year = "1976",
month = "8",
day = "1",
doi = "10.1021/bi00661a007",
language = "English",
volume = "15",
pages = "3438--3445",
journal = "Biochemistry",
issn = "0006-2960",
number = "16",

}

Formation of δ1-Acetoxytryptophan-62 in the Oxidation of Tryptophan-62 of Hen Egg-White Lysozyme by N-Bromosuccinimide in Acetate Buffer. / Norton, Raymond S.; Allerhand, Adam.

In: Biochemistry, Vol. 15, No. 16, 01.08.1976, p. 3438-3445.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Formation of δ1-Acetoxytryptophan-62 in the Oxidation of Tryptophan-62 of Hen Egg-White Lysozyme by N-Bromosuccinimide in Acetate Buffer

AU - Norton, Raymond S.

AU - Allerhand, Adam

PY - 1976/8/1

Y1 - 1976/8/1

N2 - The reaction of equimolar amounts of N-bromosuccinimide and hen egg-white lysozyme in acetate buffer, under the conditions of Hayashi et al. (Hayashi, K., Imoto, T., Funatsu, G. and Funatsu, M. (1965), J. Biochem. (Tokyo) 58, 227), yields a protein mixture that has a time-dependent 13C-NMR spectrum. The initial natural-abundance 13C-NMR spectrum indicates the presence of about equal amounts of [oxindolealanine-62]lysozyme and [δ1-acetoxytryptophan-62] lysozyme. The latter converts to [oxindolealanine-62] lysozyme w ith a half-life of about 2 days at 25 °C and pH 3.9. Two observations indicate that the source of the acetylgroup of δ1-acetoxytryptophan-62 is the acetate buffer. First, the spectrum of a lysozyme sample treated with N-bromosuccinimide in the presence of [1-13C]acetate yields a verystrong acetyl ester carbonyl resonance. The time dependence of the intensity of this resonance yields a half-life of 44 h for [δ1-acetoxytryptophan-62]lysozyme. Second, the initial natural-abundance 13C-NMR spectrum of a lysozyme sample treated with A′-bromosuccinimide in the absence of acetate indicates essentially complete conversion of tryptophan-62 into oxindolealanine.

AB - The reaction of equimolar amounts of N-bromosuccinimide and hen egg-white lysozyme in acetate buffer, under the conditions of Hayashi et al. (Hayashi, K., Imoto, T., Funatsu, G. and Funatsu, M. (1965), J. Biochem. (Tokyo) 58, 227), yields a protein mixture that has a time-dependent 13C-NMR spectrum. The initial natural-abundance 13C-NMR spectrum indicates the presence of about equal amounts of [oxindolealanine-62]lysozyme and [δ1-acetoxytryptophan-62] lysozyme. The latter converts to [oxindolealanine-62] lysozyme w ith a half-life of about 2 days at 25 °C and pH 3.9. Two observations indicate that the source of the acetylgroup of δ1-acetoxytryptophan-62 is the acetate buffer. First, the spectrum of a lysozyme sample treated with N-bromosuccinimide in the presence of [1-13C]acetate yields a verystrong acetyl ester carbonyl resonance. The time dependence of the intensity of this resonance yields a half-life of 44 h for [δ1-acetoxytryptophan-62]lysozyme. Second, the initial natural-abundance 13C-NMR spectrum of a lysozyme sample treated with A′-bromosuccinimide in the absence of acetate indicates essentially complete conversion of tryptophan-62 into oxindolealanine.

UR - http://www.scopus.com/inward/record.url?scp=0017098299&partnerID=8YFLogxK

U2 - 10.1021/bi00661a007

DO - 10.1021/bi00661a007

M3 - Article

VL - 15

SP - 3438

EP - 3445

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 16

ER -