[125I]Iododesethyl tamoxifen aziridine: Synthesis and covalent labeling of the estrogen receptor with an iodine-labeled affinity label

Francesco G. Salituro, Kathryn E. Carlson, Jonathan F. Elliston, Benita S Katzenellenbogen, John A. Katzenellenbogen

Research output: Contribution to journalArticle

Abstract

Iododesethyl tamoxifen aziridine (I-Tam-Az), an analog of the estrogen receptor-affinity label tamoxifen aziridine (Tam-Az) in which the ethyl group has been replaced by an iodine, has been prepared by two routes: (a) metallation of a bromotriarylethylene system, followed by reaction with iodine, and aziridinylation, and (b) direct iodination of a trimethylstannyl triarylethylene system that is the immediate precursor of I-Tam-Az. The latter method can be used to prepare [125I]I-Tam-Az rapidly and in good yield, both at carrier-added and no-carrier-added levels; specific activities greater than 200 Ci mmol have been obtained. In competitive radiometric binding assays with the estrogen receptor, I-Tam-Az has an apparent affinity of ca. 20%, equivalent to that of Tam-Az. It also undergoes rapid and selective time-dependent, irreversible binding to the estrogen receptor. [125I]I-Tam-Az reacts covalently with estrogen receptor in uterine cytosol preparations; its attachment is rapid and efficient, but somewhat less selective than that of Tam-Az. Estrogen receptor in intact MCF-7 human breast cancer cells can also be labeled with [125I]I-Tam-Az, and autoradiographic analysis of salt extracts of labeled nuclear estrogen receptor on SDS-polyacrylamide slab gels shows highly selective labeling of a 65K protein. [125I]I-Tam-Az is an efficient, selective affinity label for the estrogen receptor, available at high specific activity, and should be useful in studies on estrogen receptor structure, dynamics, and chromatin interactions.

Original languageEnglish (US)
Pages (from-to)287-313
Number of pages27
JournalSteroids
Volume48
Issue number5-6
DOIs
StatePublished - Jan 1 1986

Fingerprint

Affinity Labels
Iodine
Estrogen Receptors
Labeling
Competitive Binding
tamoxifen aziridine
Halogenation
Cytosol
Chromatin
Assays
Salts
Cells
Breast Neoplasms

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology
  • Pharmacology
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

[125I]Iododesethyl tamoxifen aziridine : Synthesis and covalent labeling of the estrogen receptor with an iodine-labeled affinity label. / Salituro, Francesco G.; Carlson, Kathryn E.; Elliston, Jonathan F.; Katzenellenbogen, Benita S; Katzenellenbogen, John A.

In: Steroids, Vol. 48, No. 5-6, 01.01.1986, p. 287-313.

Research output: Contribution to journalArticle

Salituro, Francesco G. ; Carlson, Kathryn E. ; Elliston, Jonathan F. ; Katzenellenbogen, Benita S ; Katzenellenbogen, John A. / [125I]Iododesethyl tamoxifen aziridine : Synthesis and covalent labeling of the estrogen receptor with an iodine-labeled affinity label. In: Steroids. 1986 ; Vol. 48, No. 5-6. pp. 287-313.
@article{a981a385b0574f53ae346df7b9895393,
title = "[125I]Iododesethyl tamoxifen aziridine: Synthesis and covalent labeling of the estrogen receptor with an iodine-labeled affinity label",
abstract = "Iododesethyl tamoxifen aziridine (I-Tam-Az), an analog of the estrogen receptor-affinity label tamoxifen aziridine (Tam-Az) in which the ethyl group has been replaced by an iodine, has been prepared by two routes: (a) metallation of a bromotriarylethylene system, followed by reaction with iodine, and aziridinylation, and (b) direct iodination of a trimethylstannyl triarylethylene system that is the immediate precursor of I-Tam-Az. The latter method can be used to prepare [125I]I-Tam-Az rapidly and in good yield, both at carrier-added and no-carrier-added levels; specific activities greater than 200 Ci mmol have been obtained. In competitive radiometric binding assays with the estrogen receptor, I-Tam-Az has an apparent affinity of ca. 20{\%}, equivalent to that of Tam-Az. It also undergoes rapid and selective time-dependent, irreversible binding to the estrogen receptor. [125I]I-Tam-Az reacts covalently with estrogen receptor in uterine cytosol preparations; its attachment is rapid and efficient, but somewhat less selective than that of Tam-Az. Estrogen receptor in intact MCF-7 human breast cancer cells can also be labeled with [125I]I-Tam-Az, and autoradiographic analysis of salt extracts of labeled nuclear estrogen receptor on SDS-polyacrylamide slab gels shows highly selective labeling of a 65K protein. [125I]I-Tam-Az is an efficient, selective affinity label for the estrogen receptor, available at high specific activity, and should be useful in studies on estrogen receptor structure, dynamics, and chromatin interactions.",
author = "Salituro, {Francesco G.} and Carlson, {Kathryn E.} and Elliston, {Jonathan F.} and Katzenellenbogen, {Benita S} and Katzenellenbogen, {John A.}",
year = "1986",
month = "1",
day = "1",
doi = "10.1016/0039-128X(86)90017-6",
language = "English (US)",
volume = "48",
pages = "287--313",
journal = "Steroids",
issn = "0039-128X",
publisher = "Elsevier Inc.",
number = "5-6",

}

TY - JOUR

T1 - [125I]Iododesethyl tamoxifen aziridine

T2 - Synthesis and covalent labeling of the estrogen receptor with an iodine-labeled affinity label

AU - Salituro, Francesco G.

AU - Carlson, Kathryn E.

AU - Elliston, Jonathan F.

AU - Katzenellenbogen, Benita S

AU - Katzenellenbogen, John A.

PY - 1986/1/1

Y1 - 1986/1/1

N2 - Iododesethyl tamoxifen aziridine (I-Tam-Az), an analog of the estrogen receptor-affinity label tamoxifen aziridine (Tam-Az) in which the ethyl group has been replaced by an iodine, has been prepared by two routes: (a) metallation of a bromotriarylethylene system, followed by reaction with iodine, and aziridinylation, and (b) direct iodination of a trimethylstannyl triarylethylene system that is the immediate precursor of I-Tam-Az. The latter method can be used to prepare [125I]I-Tam-Az rapidly and in good yield, both at carrier-added and no-carrier-added levels; specific activities greater than 200 Ci mmol have been obtained. In competitive radiometric binding assays with the estrogen receptor, I-Tam-Az has an apparent affinity of ca. 20%, equivalent to that of Tam-Az. It also undergoes rapid and selective time-dependent, irreversible binding to the estrogen receptor. [125I]I-Tam-Az reacts covalently with estrogen receptor in uterine cytosol preparations; its attachment is rapid and efficient, but somewhat less selective than that of Tam-Az. Estrogen receptor in intact MCF-7 human breast cancer cells can also be labeled with [125I]I-Tam-Az, and autoradiographic analysis of salt extracts of labeled nuclear estrogen receptor on SDS-polyacrylamide slab gels shows highly selective labeling of a 65K protein. [125I]I-Tam-Az is an efficient, selective affinity label for the estrogen receptor, available at high specific activity, and should be useful in studies on estrogen receptor structure, dynamics, and chromatin interactions.

AB - Iododesethyl tamoxifen aziridine (I-Tam-Az), an analog of the estrogen receptor-affinity label tamoxifen aziridine (Tam-Az) in which the ethyl group has been replaced by an iodine, has been prepared by two routes: (a) metallation of a bromotriarylethylene system, followed by reaction with iodine, and aziridinylation, and (b) direct iodination of a trimethylstannyl triarylethylene system that is the immediate precursor of I-Tam-Az. The latter method can be used to prepare [125I]I-Tam-Az rapidly and in good yield, both at carrier-added and no-carrier-added levels; specific activities greater than 200 Ci mmol have been obtained. In competitive radiometric binding assays with the estrogen receptor, I-Tam-Az has an apparent affinity of ca. 20%, equivalent to that of Tam-Az. It also undergoes rapid and selective time-dependent, irreversible binding to the estrogen receptor. [125I]I-Tam-Az reacts covalently with estrogen receptor in uterine cytosol preparations; its attachment is rapid and efficient, but somewhat less selective than that of Tam-Az. Estrogen receptor in intact MCF-7 human breast cancer cells can also be labeled with [125I]I-Tam-Az, and autoradiographic analysis of salt extracts of labeled nuclear estrogen receptor on SDS-polyacrylamide slab gels shows highly selective labeling of a 65K protein. [125I]I-Tam-Az is an efficient, selective affinity label for the estrogen receptor, available at high specific activity, and should be useful in studies on estrogen receptor structure, dynamics, and chromatin interactions.

UR - http://www.scopus.com/inward/record.url?scp=0022922758&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022922758&partnerID=8YFLogxK

U2 - 10.1016/0039-128X(86)90017-6

DO - 10.1016/0039-128X(86)90017-6

M3 - Article

C2 - 3445283

AN - SCOPUS:0022922758

VL - 48

SP - 287

EP - 313

JO - Steroids

JF - Steroids

SN - 0039-128X

IS - 5-6

ER -