Elsevier

Steroids

Volume 56, Issue 8, August 1991, Pages 402-410
Steroids

Size and steroid-binding characterization of membrane-associated glucocorticoid receptor in S-49 lymphoma cells

https://doi.org/10.1016/0039-128X(91)90028-TGet rights and content

Abstract

The precise mechanism for glucocorticoid-mediated lymphocytolysis is not understood, although it is presumed to be receptor mediated. We have recently presented evidence that this response is mediated by a specialized form of the glucocorticoid receptor (GR) that resides in the plasma membrane (mGR). Confirmation of the previous receptor identification studies in a population of S-49 cells enriched for mGR is now made using another antibody specific for the rodent GR, BUGR-2. The membrane resident receptor could be labeled competitively with the affinity ligand dexamethasone21-mesylate, and Scatchard analysis of whole cell binding revealed that receptor number, but not the affinity for hormone, varied between the mGR-enriched and -deficient cell populations. Steroid specificity displacement analyses showed an order of affinities as follows: triamcinolone acetonide > progesterone > dexamethasone > testosterone = estrogen. Studies of mGR by one- and two-dimensional gel electrophoresis, immunoblot, autoradiography, and density gradients revealed a species with an equivalent size to cytosolic receptor as well as multiple higher molecular weight species, confirming earlier studies. To offer a possible explanation for the nucleic acid origins of the mGR, RNA from the mGR-enriched cells was probed with rat GR cDNA; mGR-enriched cells contained higher levels of GR mRNA. Possible molecular etiologies of larger receptor species in membrane are discussed. (Steroids 56:402–410, 1991)

References (56)

  • AC Towle et al.

    Steroid binding to synaptic plasma membrane: differential binding ofglucocorticoids and gonadal steroids

    J Steroid Biochem

    (1983)
  • R Patino et al.

    Characterization of membrane receptor activity for 17α, 20β, 21-trihydroxy-4-pregnen-3-one in ovaries of spotted seatrout (Cynoscion nebulosus

    Gen Comp Endocrinol

    (1990)
  • LD Smith et al.

    The interaction of steroids with Rana pipiens oocytes in the induction of maturation

    Dev Biol

    (1971)
  • SE Sadler et al.

    Studies of a plasma membrane steroid receptor in Xenopus oocytes using the synthetic progestin RU 486

    J Steroid Biochem

    (1985)
  • HN Claman

    Corticosteroids and lymphocytolysis

    N Engt J Med

    (1972)
  • A Goldin et al.

    The chemotherapy of human and animal acute leukemia

    Cancer Chemother Rep

    (1976)
  • ME Lippman et al.

    Glucocorticoid-binding in human acute lymphoblastic leukemic cells

    J Clin Invest

    (1973)
  • ME Lippman et al.

    Clinical implications of glucocorticoid receptor in human leukemia

    Cancer Res

    (1978)
  • MR Norman et al.

    Characterization of a glucocorticoid-sensitive human lymphoid cell line

    Cancer Res

    (1977)
  • S Bourgeois et al.

    Glucocorticoid resistance in murine lymphoma and thymoma lines

    Cancer Res

    (1977)
  • D Horibato et al.

    Mouse myelomas and lymphomas in culture

    Exp Cell Res

    (1970)
  • DP Gruol et al.

    Cyclic AMP-dependent protein kinase modulation of glucocorticoid-induced cytolytic response in murine T-lymphoma cells

    Mol Endocrinol

    (1989)
  • GR Crabtree et al.

    Glucocorticoid receptors and sensitivity of isolated human leukemia and lymphoma cells

    Cancer Res

    (1978)
  • EJ Peck et al.

    Estrophylic binding sites of the uterus. Relationship to uptake and retention of estradiol in vitro

    Biochemistry

    (1973)
  • N Touchette

    Man bites dogma: a new role for steroid hormones. Steroid hormones, classical transcription regulators, may also bind to cell membrane receptors

    J NIH Res

    (1990)
  • CM Szego et al.

    Membrane recognition and effector sites in steroid hormone action

  • SY Hua et al.

    Membrane receptor-mediated electrophysiological effects of glucocorticoids on mammalian neurons

    Endocrinology

    (1989)
  • B Gametchu

    Glucocorticoid receptor-like antigen in lymphoma cell membranes: correlation to cell lysis

    Science

    (1987)
  • Cited by (46)

    • Receptor Transduction Pathways Mediating Hormone Action

      2020, Sperling Pediatric Endocrinology: Expert Consult - Online and Print
    • Glucocorticoid receptor selectively mediates stress-induced suppression of innate immunity in the House Sparrow, Passer domesticus

      2018, Developmental and Comparative Immunology
      Citation Excerpt :

      In addition, CORT can bind to plasma membrane receptors (Gametchu, 1987; Moore and Orchinik, 1994; Breuner and Orchinik, 2009; Espinoza et al., 2017). Intracellular GR can associate with the plasma membrane (reviewed in Groeneweg et al., 2012) and no membrane receptor for glucocorticoids with binding kinetics (birds, Breuner and Orchinik, 2001; Breuner and Orchinik, 2009; Schmidt et al., 2010; rats, Xiao et al., 2010; Rough-skinned Newt, Taricha granulosa, Orchinik et al., 1991) and molecular weight (mice; Gametchu et al., 1991) differing from those of classic GRs has been identified to date. We thus refer to the membrane receptor as the membrane-bound corticosteroid receptor (CR; reviewed in Groeneweg et al., 2012).

    • Further evidence for a membrane receptor that binds glucocorticoids in the rodent hypothalamus

      2016, Steroids
      Citation Excerpt :

      Specific [3H]-Cort binding in neuronal membranes in the newt was found to be about 85% of total binding, which is consistent with the higher membrane binding affinity in the newt brain [11]. Also consistent with lower-affinity glucocorticoid receptors in mammalian cell membranes, membrane binding sites for Dex in rat liver were shown to have a Kd value of approximately 400 nM [45], and studies using S-49 mouse lymphoma cells and human CCRF-CEM acute lymphoblastic leukemia cells [29,30] revealed a glucocorticoid Kd value of 239 nM for the membrane fraction compared to 19.5 nM for the cytosolic fraction [46]. Our findings here of Kd values of 30–42 nM suggest an approximately 10-fold higher affinity of glucocorticoid receptors in hypothalamic cell membranes.

    • Dissection of glucocorticoid receptor-mediated inhibition of the hypothalamic-pituitary-adrenal axis by gene targeting in mice

      2015, Frontiers in Neuroendocrinology
      Citation Excerpt :

      One mode of GR action occurs through non-genomic signaling pathways, that produce fast-acting effects in the order of seconds to a few minutes (de Kloet et al., 2008; Groeneweg et al., 2011a). This occurs through GC action on membrane bound receptors or cytoplasmic GR that activate rapid signaling of second messenger pathways (Bartholome et al., 2004; Cato et al., 2002; Gametchu et al., 1991; Spies et al., 2011; Zhang et al., 2012). The other mode is the classical GR function which occurs when activated GR translocates into the nucleus and functions as a transcription factor to modulate genomic signaling pathways, which can take a period of 15 min to several hours.

    • Receptor transduction pathways mediating hormone action

      2014, Pediatric Endocrinology: Fourth Edition
    View all citing articles on Scopus

    Present address: University of Texas Medical Branch, Galveston, Texas, 77550.

    View full text