Characterization of the chromatin acceptor sites for the avian oviduct progesterone receptor using monoclonal antibodies

Amy Goldberger, Michael Horton, Jerry Katzmann, Thomas C. Spelsberg

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Monoclonal antibodies (MAb) against the chromatin acceptor sites for the avian oviduct progesterone receptor were prepared with highly purified hen oviduct acceptor proteins reconstituted to hen DNA. Addition of the MAbs to a cell-free assay blocked progesterone receptor from chick oviduct (PRov) binding to native-like acceptor sites on nucleoacidic protein (NAP) representing a partially deproteinized chromatin, which has been shown to be enriched in these binding sites. However, the antibodies do not block PRov binding to pure DNA, nor do they affect the receptor itself. Estrogen receptor binding to NAP was not inhibited, supporting a receptor specificity of the PRov acceptor sites as reported previously from direct competition studies. These data support earlier studies showing that (1) the reconstituted PRov acceptor sites resemble the native sites, (2) the acceptor sites are receptor specific, and (3) the PRov binding sites of NAP are different from those of pure DNA. While some animal-species specificity in the PRov binding inhibition was observed, no tissue specificity was seen. Direct binding of the antibodies to native acceptor sites was demonstrated in an enzyme-linked immunosorbent assay (ELISA) system. The antibodies showed little recognition of free acceptor protein or DNA alone, indicating specificity for the protein-DNA complex. A partial evolutionary conservation of the nuclear acceptor sites for PRov was shown by the fact that about 50% of the inhibition seen with hen NAP was obtained with NAPs from several other species, and this partial cross-reactivity of the MAbs with the same NAPs from other animal species was also seen in the ELISA.

Original languageEnglish (US)
Pages (from-to)5811-5816
Number of pages6
JournalBiochemistry
Volume26
Issue number18
StatePublished - 1987
Externally publishedYes

Fingerprint

Oviducts
Progesterone Receptors
Chromatin
Monoclonal Antibodies
Proteins
DNA
Assays
Immunosorbents
Antibodies
Animals
Enzyme-Linked Immunosorbent Assay
Binding Sites
Species Specificity
Organ Specificity
Enzymes
Estrogen Receptors
Conservation
Tissue

ASJC Scopus subject areas

  • Biochemistry

Cite this

Goldberger, A., Horton, M., Katzmann, J., & Spelsberg, T. C. (1987). Characterization of the chromatin acceptor sites for the avian oviduct progesterone receptor using monoclonal antibodies. Biochemistry, 26(18), 5811-5816.

Characterization of the chromatin acceptor sites for the avian oviduct progesterone receptor using monoclonal antibodies. / Goldberger, Amy; Horton, Michael; Katzmann, Jerry; Spelsberg, Thomas C.

In: Biochemistry, Vol. 26, No. 18, 1987, p. 5811-5816.

Research output: Contribution to journalArticle

Goldberger, A, Horton, M, Katzmann, J & Spelsberg, TC 1987, 'Characterization of the chromatin acceptor sites for the avian oviduct progesterone receptor using monoclonal antibodies', Biochemistry, vol. 26, no. 18, pp. 5811-5816.
Goldberger, Amy ; Horton, Michael ; Katzmann, Jerry ; Spelsberg, Thomas C. / Characterization of the chromatin acceptor sites for the avian oviduct progesterone receptor using monoclonal antibodies. In: Biochemistry. 1987 ; Vol. 26, No. 18. pp. 5811-5816.
@article{3577ce69003748f5b893f047c3e7e681,
title = "Characterization of the chromatin acceptor sites for the avian oviduct progesterone receptor using monoclonal antibodies",
abstract = "Monoclonal antibodies (MAb) against the chromatin acceptor sites for the avian oviduct progesterone receptor were prepared with highly purified hen oviduct acceptor proteins reconstituted to hen DNA. Addition of the MAbs to a cell-free assay blocked progesterone receptor from chick oviduct (PRov) binding to native-like acceptor sites on nucleoacidic protein (NAP) representing a partially deproteinized chromatin, which has been shown to be enriched in these binding sites. However, the antibodies do not block PRov binding to pure DNA, nor do they affect the receptor itself. Estrogen receptor binding to NAP was not inhibited, supporting a receptor specificity of the PRov acceptor sites as reported previously from direct competition studies. These data support earlier studies showing that (1) the reconstituted PRov acceptor sites resemble the native sites, (2) the acceptor sites are receptor specific, and (3) the PRov binding sites of NAP are different from those of pure DNA. While some animal-species specificity in the PRov binding inhibition was observed, no tissue specificity was seen. Direct binding of the antibodies to native acceptor sites was demonstrated in an enzyme-linked immunosorbent assay (ELISA) system. The antibodies showed little recognition of free acceptor protein or DNA alone, indicating specificity for the protein-DNA complex. A partial evolutionary conservation of the nuclear acceptor sites for PRov was shown by the fact that about 50{\%} of the inhibition seen with hen NAP was obtained with NAPs from several other species, and this partial cross-reactivity of the MAbs with the same NAPs from other animal species was also seen in the ELISA.",
author = "Amy Goldberger and Michael Horton and Jerry Katzmann and Spelsberg, {Thomas C.}",
year = "1987",
language = "English (US)",
volume = "26",
pages = "5811--5816",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "18",

}

TY - JOUR

T1 - Characterization of the chromatin acceptor sites for the avian oviduct progesterone receptor using monoclonal antibodies

AU - Goldberger, Amy

AU - Horton, Michael

AU - Katzmann, Jerry

AU - Spelsberg, Thomas C.

PY - 1987

Y1 - 1987

N2 - Monoclonal antibodies (MAb) against the chromatin acceptor sites for the avian oviduct progesterone receptor were prepared with highly purified hen oviduct acceptor proteins reconstituted to hen DNA. Addition of the MAbs to a cell-free assay blocked progesterone receptor from chick oviduct (PRov) binding to native-like acceptor sites on nucleoacidic protein (NAP) representing a partially deproteinized chromatin, which has been shown to be enriched in these binding sites. However, the antibodies do not block PRov binding to pure DNA, nor do they affect the receptor itself. Estrogen receptor binding to NAP was not inhibited, supporting a receptor specificity of the PRov acceptor sites as reported previously from direct competition studies. These data support earlier studies showing that (1) the reconstituted PRov acceptor sites resemble the native sites, (2) the acceptor sites are receptor specific, and (3) the PRov binding sites of NAP are different from those of pure DNA. While some animal-species specificity in the PRov binding inhibition was observed, no tissue specificity was seen. Direct binding of the antibodies to native acceptor sites was demonstrated in an enzyme-linked immunosorbent assay (ELISA) system. The antibodies showed little recognition of free acceptor protein or DNA alone, indicating specificity for the protein-DNA complex. A partial evolutionary conservation of the nuclear acceptor sites for PRov was shown by the fact that about 50% of the inhibition seen with hen NAP was obtained with NAPs from several other species, and this partial cross-reactivity of the MAbs with the same NAPs from other animal species was also seen in the ELISA.

AB - Monoclonal antibodies (MAb) against the chromatin acceptor sites for the avian oviduct progesterone receptor were prepared with highly purified hen oviduct acceptor proteins reconstituted to hen DNA. Addition of the MAbs to a cell-free assay blocked progesterone receptor from chick oviduct (PRov) binding to native-like acceptor sites on nucleoacidic protein (NAP) representing a partially deproteinized chromatin, which has been shown to be enriched in these binding sites. However, the antibodies do not block PRov binding to pure DNA, nor do they affect the receptor itself. Estrogen receptor binding to NAP was not inhibited, supporting a receptor specificity of the PRov acceptor sites as reported previously from direct competition studies. These data support earlier studies showing that (1) the reconstituted PRov acceptor sites resemble the native sites, (2) the acceptor sites are receptor specific, and (3) the PRov binding sites of NAP are different from those of pure DNA. While some animal-species specificity in the PRov binding inhibition was observed, no tissue specificity was seen. Direct binding of the antibodies to native acceptor sites was demonstrated in an enzyme-linked immunosorbent assay (ELISA) system. The antibodies showed little recognition of free acceptor protein or DNA alone, indicating specificity for the protein-DNA complex. A partial evolutionary conservation of the nuclear acceptor sites for PRov was shown by the fact that about 50% of the inhibition seen with hen NAP was obtained with NAPs from several other species, and this partial cross-reactivity of the MAbs with the same NAPs from other animal species was also seen in the ELISA.

UR - http://www.scopus.com/inward/record.url?scp=0023648327&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023648327&partnerID=8YFLogxK

M3 - Article

C2 - 3676291

AN - SCOPUS:0023648327

VL - 26

SP - 5811

EP - 5816

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 18

ER -