Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan

William A. Frazier, Ruth A. Hogue-Angeletti, Randolph Sherman, Ralph A. Bradshaw

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

The chemical reactivity and degree of solvent exposure of the two tyrosine and three tryptophan residues of mouse 2.5S nerve growth factor (NGF) have been investigated. Spectrophotometric titrations and reaction with tetranitromethane indicate that both tyrosines are solvent available to a limited extent and that neither is required for activity. While the two tyrosines appear equivalent, the tryptophans exhibit a spectrum of reactivity. The reaction of these residues with N-bromosuccinimide is a kinetically ordered process. The most rapidly reacting residue is also solvent available by the criterion of N-methylnicotinamide chloride titration and is not required for activity. Reaction of the intermediate residue destroys both biological and immunological activity, while oxidation of the third residue leads to dissociation of the 2.5S dimer. Reaction of native NGF with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide leads to modification of Trp-21 and Trp-99 and leaves Trp-76 unmodified. Modification of the rapidly oxidized residue with N-bromosuccinimide followed by reaction with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide in 6 M guanidine hydrochloride reveals the modification of Trp-99 and Trp-76, identifying Trp-21 as the solvent available residue. Computer protein modeling studies show that this chemical description of the topography of the tyrosine and tryptophan residues of NGF is in extremely good agreement with topography of the corresponding residues in the three-dimensional structure of insulin. Thus the observed similarities in the function and mechanism of action of NGF and insulin may result from regions of similar primary structure dictating related three-dimensional conformation.

Original languageEnglish (US)
Pages (from-to)3281-3293
Number of pages13
JournalBiochemistry
Volume12
Issue number17
StatePublished - 1973
Externally publishedYes

Fingerprint

Tryptophan
Topography
Tyrosine
Nerve Growth Factor
Bromosuccinimide
Titration
Tetranitromethane
Insulin
Chemical reactivity
Guanidine
Dimers
Conformations
Chlorides
Oxidation
2.5S nerve growth factor
Proteins
dimethyl(2-hydroxy-5-nitrobenzyl)sulfonium

ASJC Scopus subject areas

  • Biochemistry

Cite this

Frazier, W. A., Hogue-Angeletti, R. A., Sherman, R., & Bradshaw, R. A. (1973). Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan. Biochemistry, 12(17), 3281-3293.

Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan. / Frazier, William A.; Hogue-Angeletti, Ruth A.; Sherman, Randolph; Bradshaw, Ralph A.

In: Biochemistry, Vol. 12, No. 17, 1973, p. 3281-3293.

Research output: Contribution to journalArticle

Frazier, WA, Hogue-Angeletti, RA, Sherman, R & Bradshaw, RA 1973, 'Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan', Biochemistry, vol. 12, no. 17, pp. 3281-3293.
Frazier WA, Hogue-Angeletti RA, Sherman R, Bradshaw RA. Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan. Biochemistry. 1973;12(17):3281-3293.
Frazier, William A. ; Hogue-Angeletti, Ruth A. ; Sherman, Randolph ; Bradshaw, Ralph A. / Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan. In: Biochemistry. 1973 ; Vol. 12, No. 17. pp. 3281-3293.
@article{81c475bf9f2d4af49f22deca9bee5743,
title = "Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan",
abstract = "The chemical reactivity and degree of solvent exposure of the two tyrosine and three tryptophan residues of mouse 2.5S nerve growth factor (NGF) have been investigated. Spectrophotometric titrations and reaction with tetranitromethane indicate that both tyrosines are solvent available to a limited extent and that neither is required for activity. While the two tyrosines appear equivalent, the tryptophans exhibit a spectrum of reactivity. The reaction of these residues with N-bromosuccinimide is a kinetically ordered process. The most rapidly reacting residue is also solvent available by the criterion of N-methylnicotinamide chloride titration and is not required for activity. Reaction of the intermediate residue destroys both biological and immunological activity, while oxidation of the third residue leads to dissociation of the 2.5S dimer. Reaction of native NGF with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide leads to modification of Trp-21 and Trp-99 and leaves Trp-76 unmodified. Modification of the rapidly oxidized residue with N-bromosuccinimide followed by reaction with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide in 6 M guanidine hydrochloride reveals the modification of Trp-99 and Trp-76, identifying Trp-21 as the solvent available residue. Computer protein modeling studies show that this chemical description of the topography of the tyrosine and tryptophan residues of NGF is in extremely good agreement with topography of the corresponding residues in the three-dimensional structure of insulin. Thus the observed similarities in the function and mechanism of action of NGF and insulin may result from regions of similar primary structure dictating related three-dimensional conformation.",
author = "Frazier, {William A.} and Hogue-Angeletti, {Ruth A.} and Randolph Sherman and Bradshaw, {Ralph A.}",
year = "1973",
language = "English (US)",
volume = "12",
pages = "3281--3293",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "17",

}

TY - JOUR

T1 - Topography of mouse 2.5S nerve growth factor. Reactivity of tyrosine and tryptophan

AU - Frazier, William A.

AU - Hogue-Angeletti, Ruth A.

AU - Sherman, Randolph

AU - Bradshaw, Ralph A.

PY - 1973

Y1 - 1973

N2 - The chemical reactivity and degree of solvent exposure of the two tyrosine and three tryptophan residues of mouse 2.5S nerve growth factor (NGF) have been investigated. Spectrophotometric titrations and reaction with tetranitromethane indicate that both tyrosines are solvent available to a limited extent and that neither is required for activity. While the two tyrosines appear equivalent, the tryptophans exhibit a spectrum of reactivity. The reaction of these residues with N-bromosuccinimide is a kinetically ordered process. The most rapidly reacting residue is also solvent available by the criterion of N-methylnicotinamide chloride titration and is not required for activity. Reaction of the intermediate residue destroys both biological and immunological activity, while oxidation of the third residue leads to dissociation of the 2.5S dimer. Reaction of native NGF with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide leads to modification of Trp-21 and Trp-99 and leaves Trp-76 unmodified. Modification of the rapidly oxidized residue with N-bromosuccinimide followed by reaction with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide in 6 M guanidine hydrochloride reveals the modification of Trp-99 and Trp-76, identifying Trp-21 as the solvent available residue. Computer protein modeling studies show that this chemical description of the topography of the tyrosine and tryptophan residues of NGF is in extremely good agreement with topography of the corresponding residues in the three-dimensional structure of insulin. Thus the observed similarities in the function and mechanism of action of NGF and insulin may result from regions of similar primary structure dictating related three-dimensional conformation.

AB - The chemical reactivity and degree of solvent exposure of the two tyrosine and three tryptophan residues of mouse 2.5S nerve growth factor (NGF) have been investigated. Spectrophotometric titrations and reaction with tetranitromethane indicate that both tyrosines are solvent available to a limited extent and that neither is required for activity. While the two tyrosines appear equivalent, the tryptophans exhibit a spectrum of reactivity. The reaction of these residues with N-bromosuccinimide is a kinetically ordered process. The most rapidly reacting residue is also solvent available by the criterion of N-methylnicotinamide chloride titration and is not required for activity. Reaction of the intermediate residue destroys both biological and immunological activity, while oxidation of the third residue leads to dissociation of the 2.5S dimer. Reaction of native NGF with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide leads to modification of Trp-21 and Trp-99 and leaves Trp-76 unmodified. Modification of the rapidly oxidized residue with N-bromosuccinimide followed by reaction with dimethyl-(2-hydroxy-5-nitrobenzyl)sulfonium bromide in 6 M guanidine hydrochloride reveals the modification of Trp-99 and Trp-76, identifying Trp-21 as the solvent available residue. Computer protein modeling studies show that this chemical description of the topography of the tyrosine and tryptophan residues of NGF is in extremely good agreement with topography of the corresponding residues in the three-dimensional structure of insulin. Thus the observed similarities in the function and mechanism of action of NGF and insulin may result from regions of similar primary structure dictating related three-dimensional conformation.

UR - http://www.scopus.com/inward/record.url?scp=0015859963&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0015859963&partnerID=8YFLogxK

M3 - Article

VL - 12

SP - 3281

EP - 3293

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 17

ER -