Über Hormon-„Receptoren“ Die oestrogenbindenden Prinzipien der Erfolgsorgane

Jungblut, P. W.; Hätzel, I.; DeSombre, Eugene R.; Jensen, Elwood V.

doi:10.1007/978-3-642-88715-4_3

Cited by 12 publications

(4 citation statements)

References 8 publications

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“…Since the earliest application of affinity chromatography for the purification of estrogen receptors (9,19), the use of this technique has been limited by the surprising resistance of the bound receptor to elution under conditions compatible with its stability. Attachment of the steroid to the supporting matrix by an extremely long molecular bridge has been reported to facilitate release of the bound receptor (20,21) but, unless removed by chromatography over heparin-Sepharose (18), enzymes present in target tissue extracts tend to cleave the ester or amide groups in the macromolecular spacer linking the steroid to the supporting medium.…”

Section: Discussionmentioning

confidence: 99%

Monoclonal antibodies to human estrogen receptor.

Greene¹,

Nolan²,

Engler³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

446

110

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Extranuclear estrogen receptor protein (estrophilin) of MCF-7 human breast cancer celIs was purified by passage of the cytosol fraction of a cell homogenate through an affinity column of estradiol linked to Sepharose by a substituted di-n-propyl sulfide bridge in the 17a position. Elution The preparation of specific antibodies to estrogen receptors has provided a means of recognizing receptor proteins other than by their binding to labeled steroid. Antibodies to calf uterine estrophilin,-generated in either rabbits (1-3) or a goat (3, 4), crossreact with estrogen receptors, but not with androgen or progesterone receptors, from a wide variety of animal species. The hormone specificity and crossreactivity of the rabbit and goat antiestrophilins make them attractive as probes for the study of receptor structure and function, but their use in immunochemical procedures for receptor purification, assay, and cellular localization is limited by the heterogeneity of these antibody preparations. Monoclonal antiestrophilin antibodies secreted by hybridoma cells, obtained by fusion of mouse myeloma cells with splenic lymphocytes from a Lewis rat immunized with calf uterine estrophilin, have been described (4, 5) but, like the immunoglobulin in the serum of the parent rat, they reacted specifically with calf estrophilin and thus are unsuitable for the study of receptors from species other than calf.This Preparation of Estradiol Affinity Adsorbent. 17a-Allylestradiol 3-acetate, prepared by the reaction of estrone with allylmagnesium chloride (9) followed by acetylation of the phenolic group (10), was converted to the side-chain epoxide by treatment with m-chloroperoxybenzoic acid (11). Reaction of the epoxide with reduced thiopropyl-Sepharose (2-hyAbbreviations: E*R, [3H]estradiol-estrophilin complex; i-Ig, immunoglobulin secreted by hybridomas derived from immunized rat; n-Ig, immunoglobulin from serum of nonimmunized rat; HAT, hypoxanthine/amethopterin/thymidine; P3, P3-X63-Ag8; Sp2/0, Sp2/0-Agl4. 5115The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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Section: Discussionmentioning

confidence: 99%

Monoclonal antibodies to human estrogen receptor.

Greene¹,

Nolan²,

Engler³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

446

110

View full text Add to dashboard Cite

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“…This property is bestowed by the receptor, which binds the hormone non-covalently but with high affinity (Jungblut et al 1967). The stability of the estradiol-receptor bond at neutral pH and low temperature allowed for subcellular distribution studies by differential centrifugation of homogenates, recognizing the nucleus as the prominent site of bound estradiol.…”

Section: Introductionmentioning

confidence: 99%

Identification of target cells by immunohistochemical detection of covalently rearranged estradiol in rehydrated paraffin sections

Jungblut

Sierralta

1998

Histochemistry and Cell Biology

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“…Extracts of calf Uterus nuclei with buffered 0.3 mol/1 KC1 were found to* contain a 5 S receptor molecule (15,16). First attempts at interconversion by a lengthy sah exchange through Sephadex columns failed (16,17), while the addition of salt to freshly prepared low ionic strength cytosol gave rise to a 4-5 S receptor peak in density gradient centrifugation (18). A still growing host of papers has since been devoted to this problem and its likely physiological implications.…”

Section: The 8-10 "Storage" Form Of "Cytosol" Oestradiol Receptormentioning

confidence: 98%

Subcellular Distribution, Properties and Interrelationship of Oestrogen Receptors in Endometrium and Other Target Tissues

Jungblut¹,

Hekim²,

Meyer³

et al. 1983

Clinical Chemistry and Laboratory Medicine

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More than half of the extranuclear receptor content of resting cells is associated with cytoplasmic structures. Subfractionation of mierosomes reveals a preponderance of "basic" (low electrophoretic mobility) receptor in rough endoplasmic reticulum. Surfynol-dithiothreitol extracts of smooth membranes are rieh in "acidic" (high electrophoretic mobility) receptor. Trypsin increases the yields up to seven-fold, and this increase is correlated (r = 0.993) with the acidic receptor content and S'^nucleotidase activity of these microsomal preparations. Acidic microsomal, "cytosolic" and nuclear receptor can be degraded to the "basic" variety by streptomyces hyaluronidase. All forms give rise to a tryptic fragment with unchanged affinity for oestradiol and dimerization ability. Basic receptor isolated after enzymatic conversion of acidic receptor is microheterogenous on isoelectric focüsing, but gives rise to only one precipitation arc versus the IgG fraction of a polyclonal antiserum. The precipitation arc can be recharged with labelled oestradiol and autoradiographed. Non-immune IgG's form (unspecific) soluble complexes with oestrogen receptors, but not with their tryptic fragments. The polyclonal antioestrogen receptor IgG fraction precipitates the oestradiol-tagged tryptic receptor fragment from all subcellular sources of all homologous (porcine) and heterologous (human, ovine, bovine, goat, rabbit, guinea pig, rat) target tissues tested. Organ specificity can therefore be excluded and a high degree of phylogenetic conservation of the oestrogen receptor's protein moiety is implied. The presence, in the immune IgG fraction, of steroid releasing antibodies, which apparently distort the binding site, should spur the search for monoclonal antibodies with similar properties. Subzelluläre Verteilung, Eigenschaften und Verwandtschaft der östrogenrezeptoren in Endometrium und anderen ZielorganenZusammenfassung: Mehr als die Hälfte des extranuklearen Rezeptorgehalts von ruhenden Zellen ist mit zytoplasmatischen Strukturen assoziiert. Nach Subfraktionierung von Mikrosomen wird in rauhem Ergastoplasma bevorzugt "basischer" (niedrige elektrophoretische Beweglichkeit) Rezeptor gefunden. Surfynol/Dithiothreitöl-Extrakte von glatten Membranen sind reich an "saurem" (hohe elektrophoretische Beweglichkeit) Rezeptor. Trypsin erhöht die Extraktionsausbeuten bis zum Siebenfachen in Korrelation (r = 0,993) zum Gehalt der Strukturen an saurem Rezeptor und ihrer 5 f -Nucleotidaseaktivität. Saurer Rezeptor aus Cytosol, Mikrosomen und Kernen wird durch Streptomyces-Hyaluronidase zur basischen Form abgebaut. Trypsin führt sowohl sauren als auch basischen Rezeptor in ein Fragment über, das eine unveränderte Affinität für Östradiol hat und noch dimerisieren kann. Durch enzymatische Umwandlung von saurem Rezeptor und anschließende Reinigung erhaltener basischer Rezeptor ist mikroheterogen in der isoelektrischen Fokussierung, bildet jedoch nur einen Präzipitationsbogen mit der IgG-Fraktion eines polyklonalen Antiserums. Das Immunpräzipitat kann ...

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Über Hormon-„Receptoren“ Die oestrogenbindenden Prinzipien der Erfolgsorgane

Cited by 12 publications

References 8 publications

Monoclonal antibodies to human estrogen receptor.

Monoclonal antibodies to human estrogen receptor.

Identification of target cells by immunohistochemical detection of covalently rearranged estradiol in rehydrated paraffin sections

Subcellular Distribution, Properties and Interrelationship of Oestrogen Receptors in Endometrium and Other Target Tissues

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