Abstract:
:Most secreted proteins are modified post-translationally with the addition of carbohydrate. It has been difficult to use crystallography to solve the structures of these proteins due to the inherent heterogeneity of the carbohydrate. The structure of the chemically deglycosylated form (hydrogen fluoride treated) of human chorionic gonadotropin (hCG) has been solved through crystallographic techniques. Unfortunately this form of hCG is not biologically active, and exhibits immunochemical differences from native hormone. In addition, subunit interactions appear altered after chemical deglycosylation as indicated by the increased thermal stability of the HF-treated hormone. The Asn 52 glycan on the alpha-subunit of hCG has been identified as being required for biological activity, it is, therefore, of physiological importance to determine the structure of the hormone with its carbohydrate intact. Also, it has not been possible to obtain crystals of the individual glycosylated subunits of hCG. Therefore an alternative method to solve the structure of the biologically active form of the hormone in solution as well as its separated subunits is necessary. Structural information utilizing NMR techniques can be obtained from native hCG subunits in solution if they can be uniformly labeled with 13C and 15N isotopes. We have developed a universal nonradioactive isotope, labeling medium enriched in 13C and 15N which can be used to express uniformly labeled hCG from Chinese hamster ovary cells suitable for solving the structure of the individual subunits and ultimately that of the native, biologically active hormone. The isotopically labeled recombinant hCG and its purified subunits are essentially identical to urinary hCG on comparison by biochemical, immunochemical, biological activity and the ability of the isolated subunits to recombine to form a biologically active dimer. Mass spectrometric analysis and preliminary structural NMR data indicate that the labeling is uniform and there is greater than 90% incorporation, sufficient for complete structural determination studies. This labeled growth medium represents a technological advance which will enable the rapid solution of the structures of the other glycoprotein hormones, as well as other glycoproteins which have proven unsuitable for crystallographic study.
journal_name
Mol Cell Endocrinoljournal_title
Molecular and cellular endocrinologyauthors
Lustbader JW,Pollak S,Lobel L,Trakht I,Homans S,Brown JM,Canfield REdoi
10.1016/s0303-7207(96)03952-4subject
Has Abstractpub_date
1996-12-20 00:00:00pages
21-31issue
1-2eissn
0303-7207issn
1872-8057journal_volume
125pub_type
杂志文章,评审abstract::Our objective was to investigate the mechanisms by which the endogenous CRHR2 in white adipose tissue (WAT) regulates metabolic activities associated with lipogenesis and lipolysis under continuous exposure to hypoxia. We found that hypobaric hypoxia at a simulated altitude of 5000 m significantly reduced the body wei...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/j.mce.2014.05.012
更新日期:2014-07-05 00:00:00
abstract::Bovine adrenocortical cells were transplanted into scid mice, using a small cylinder inserted beneath the kidney capsule. The tissue formed from primary bovine adrenocortical cells replaced the essential functions of the animals' own adrenal glands, which were removed during the cell transplantation procedure. Most ad...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/s0303-7207(99)00070-2
更新日期:1999-07-20 00:00:00
abstract::Hypothalamic astrocytes secrete TGF-beta and 3 alpha,5 alpha-tetrahydro progesterone (3 alpha,5 alpha-THP) in culture. When the astrocyte-conditioned medium (ACM) was incubated with the hypothalamic cell line GT1-7, it resulted in the secretion of GnRH. Immunoneutralization with TGF-beta antibody or ultra-filteration ...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2005.11.017
更新日期:2006-02-26 00:00:00
abstract::Somatostatin has been discovered as a somatotroph release inhibitory factor (SRIF), and, indeed, it has been demonstrated that SRIF and its analogs can inhibit pituitary tumor hormone secretion and control neoplastic bulk. Several in vitro studies have contributed to the current knowledge of the mechanisms by which SR...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2007.12.010
更新日期:2008-05-14 00:00:00
abstract::We recently cloned and sequenced two kilobases of the upstream flanking region of the mouse tissue transglutaminase gene. Transfection experiments showed that this region of the transglutaminase flanking sequence was sufficient to mediate a 4-fold induction in reporter gene expression by retinoic acid. The goal of the...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(96)03826-9
更新日期:1996-07-01 00:00:00
abstract::In order to study the control of vasopressin-release, the effect of a series of potential agents was studied in an in vitro perifusion system of rat neurohypophysis after in vivo treatment with nialamide, a monoamine oxidase inhibitor. In this system, metlatonin stimulated vasopressin-release in a dose-dependent manne...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(79)90103-5
更新日期:1979-05-01 00:00:00
abstract::The number of pituitary cells, their size, hormonal content and release and response to external cues varies between day and night and during the estrus cycle. Previous studies have demonstrated that pituitary cells proliferate rhythmically and that estradiol (E(2)) is a mitogen of alpha T3 cells. We, therefore, studi...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/s0303-7207(03)00119-9
更新日期:2003-05-30 00:00:00
abstract::1,25-Dihydroxyvitamin D (1,25(OH)(2)D) plays a critical role in calcium and phosphorus (Pi) metabolism, bone growth, and tissue differentiation. The synthesis of 1,25(OH)(2)D in the proximal renal tubule is the primary determinant of its circulating concentration and is mediated by the mitochondrial enzyme, 25-hydroxy...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2011.08.030
更新日期:2011-12-05 00:00:00
abstract::Regulation of hormonal, insulin/IGF-1 (Ins/IGF-1) signaling activities, and pathways of the intrinsic generation of reactive oxygen species (ROS) play a role in aging and longevity determination. In this review we discuss the cross-talk between these pathways as mechanisms of signaling that may be important factors in...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2008.11.025
更新日期:2009-02-05 00:00:00
abstract::The aim of this study was to investigate whether co-culture of human islets with adipose-derived stem cells (ASCs) can improve islet quality and to evaluate which factors play a role in the protective effect of ASCs against islet dysfunction. Islets and ASCs were cultured in three experimental groups for 24 h, 48 h, a...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/j.mce.2020.110729
更新日期:2020-04-05 00:00:00
abstract::The immune system and the neuroendocrine system are functionally interactive. Lymphocytes possess opioid receptors as well as growth hormone receptors (GHR) by which opioids and growth hormone (GH) may modulate immune functions. In this study, we have investigated the effects of morphine on [125I]hGH binding and GHR g...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/s0303-7207(97)00197-4
更新日期:1997-12-12 00:00:00
abstract::In cultured human lymphocytes (IM-9) and in isolated rat adipocytes human growth hormone is substrate for a receptor-mediated degradation. When the cells are incubated with monoiodinated human growth hormone half of the radioactivity dissociating from the cells is in the form of [125I]monoiodotyrosine. Since IM-9 lymp...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(85)90145-5
更新日期:1985-06-01 00:00:00
abstract::Steroidogenesis refers to the de novo synthesis of steroid hormones from cholesterol by a number of sequential enzyme catalysed reactions in the adrenal and the gonads. In addition, circulating steroid hormone precursors are further metabolised in selected peripheral tissues. It has been suggested that the biosynthesi...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2016.09.004
更新日期:2017-02-05 00:00:00
abstract::Steroidogenic Factor 1 (SF-1, Ad4bp, NR5A1) is a nuclear receptor expressed mainly in the adrenals and gonads. It activates the transcription of genes in steroidogenesis, reproduction, and energy metabolism. In addition, it also regulates the growth and differentiation of adrenogonadal primodial cells. SF-1 resides in...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2012.11.019
更新日期:2013-05-22 00:00:00
abstract::Type 1 diabetes (T1D) is an autoimmune disease resulting from the loss of pancreatic β cells and subsequent insulin production. Novel interventional therapies are urgently needed that can protect existing β cells from cytokine-induced death and enhance their function before symptomatic onset. Our initial evidence is s...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/j.mce.2019.110491
更新日期:2019-08-20 00:00:00
abstract::The high-density lipoprotein (HDL) pathway in rat adrenocortical cells was studied at the electron microscopic level in vitro via colloidal gold labelling. Steroid hormone assays were performed to confirm that the cells remained intact, viable, responsive to ACTH under the applied conditions, and to reveal the steroid...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(86)90062-6
更新日期:1986-02-01 00:00:00
abstract::An additional form of basic fibroblast growth factor (FGF), possessing full biological activity, has been isolated from bovine pituitary extracts. This polypeptide mitogen exhibits an apparent molecular weight of 17,500 by SDS-PAGE and lacks the first four N-terminal amino acid residues (Pro-Ala-Leu-Pro) of a previous...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(87)90028-1
更新日期:1987-06-01 00:00:00
abstract::1,25(OH)2D3 (vitamin D) is well-recognized as a neurosteroid that modulates multiple brain functions. A growing body of evidence indicates that vitamin D plays a pivotal role in brain development, neurotransmission, neuroprotection and immunomodulation. However, the precise molecular mechanisms by which vitamin D exer...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2017.05.035
更新日期:2017-09-15 00:00:00
abstract::This study examines the effect of preincubation of sheep ovarian follicles with hCG or FSH on the ability of isolated theca and granulosa to increase cAMP levels in response to a second incubation with hCG or FSH. Secondly, the effect of preincubation of small follicles with hCG on androstenedione and testosterone pro...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(79)90083-2
更新日期:1979-03-01 00:00:00
abstract::The expression of genes encoding inhibin/activin subunits and activin receptor was examined in four cultured Leydig tumor cells (MA-10, I-10, R2C, and LC-540). Inhibin alpha-subunit gene was highly expressed in Leydig tumor cell lines except LC-540. Both inhibin beta-A- and beta-B-subunit mRNAs were present in low lev...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(93)90061-n
更新日期:1993-07-01 00:00:00
abstract::The melanocortin receptor family consists of 5 members which belong to the GPCR superfamily. Their specific ligands, the melanocortins are peptide hormones which are formed by the proteolytic cleavage of the proopiomelanocortin (POMC) protein. It is now recognised that certain GPCRs require accessory proteins for thei...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2010.07.015
更新日期:2011-01-15 00:00:00
abstract::The involvement of continuous protein synthesis in the mechanisms of crustacean steroidogenesis was investigated using crayfish molting glands (Y-organs). During intermolt, Y-organ steroidogenic activity is low. Eyestalk ablation initiates premolt which is characterized by a rapid increase in the production of ecdyste...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(95)03489-t
更新日期:1995-03-01 00:00:00
abstract::The template for our understanding of the physiological role of the adrenal cortex was set by Hans Selye, who demonstrated its key involvement in the response to stress, of whatever origin, and who also introduced the terms glucocorticoid and mineralocorticoid. Despite this, from the late 1940s on there was certainly ...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2008.09.008
更新日期:2009-03-05 00:00:00
abstract::The detergent-soluble extract of rat ovary plasma membranes contained a Gs protein of about 100 kDa as shown by its elution behavior on a Bio Gel A-1.5m column. However, the cell membranes exposed to hCG (37 degrees C, 15 min) contained in addition a higher molecular weight Gs protein complex of 300 kDa comprised of h...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(91)90147-k
更新日期:1991-09-01 00:00:00
abstract::Pro-opiomelanocortin (POMC) is a prohormone which undergoes extensive tissue and cell specific post-translational processing producing a number of active peptides with diverse biological roles ranging from control of adrenal function to pigmentation to the regulation of feeding. One approach to unraveling the complexi...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/j.mce.2010.12.032
更新日期:2011-04-10 00:00:00
abstract::The Nile tilapia, Oreochromis niloticus, is a gonochoristic teleost fish with an XX/XY genetic system and is an excellent model for gonadal sex differentiation. In the present study, we screened novel genes that were expressed predominantly in either XY or XX undifferentiated gonads during the critical period for diff...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/j.mce.2015.08.008
更新日期:2015-11-05 00:00:00
abstract::We have produced transgenic (TG) mice expressing under the ubiquitin C promoter either the glycoprotein hormone common alpha-subunit (C(alpha)) or human chorionic gonadotropin (hCG) beta-subunit. C(alpha) overexpression alone had no phenotypic effect, but the hCG(beta) expressing females, presenting with moderately el...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章,评审
doi:10.1016/j.mce.2004.10.013
更新日期:2005-04-29 00:00:00
abstract::Glucocorticoid hormones suppress circulating concentrations of thyrotrophin (TSH), but their effect on synthesis of TSH in the pituitary gland is unclear. We have examined the influence of the glucocorticoid dexamethasone on serum TSH, pituitary TSH content and TSH beta- and alpha-subunit mRNA concentrations in pituit...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(89)90064-6
更新日期:1989-06-01 00:00:00
abstract::1,25-Dihydroxyvitamin D(3) (1,25D(3)), the active metabolite of vitamin D(3), inhibits breast cancer cell growth in vivo and in vitro. To examine mechanisms of 1,25D(3) induced growth arrest and apoptosis, cell lines were established from DMBA induced mammary tumors derived from vitamin D(3) receptor knockout (VDRKO) ...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/s0303-7207(02)00416-1
更新日期:2003-02-28 00:00:00
abstract::Insulin-like growth factor-I (IGF-I) is involved in the growth and development of liver and brain during fetal life by acting through specific plasma membrane receptors. In an attempt to determine how the changes in IGF-I receptor number are regulated during development, we have compared [125I]IGF-I binding to membran...
journal_title:Molecular and cellular endocrinology
pub_type: 杂志文章
doi:10.1016/0303-7207(94)90222-4
更新日期:1994-05-01 00:00:00