A seminar on,hormon receptor

this seminar mainly gives the information about hormonal receptor.

1. By, Sanket J. Gandhi M. PHARM (I st SEM ) Department of Pharmacology R.C. Patel Institute Of Pharmaceutical education and Research Hormonal RECEPTORs

2. Index Introduction to Hormone 3,4 Slide No. Hormonal receptor 5,6 Cell surface receptor 7-16 Nuclear receptor 17-29 Cell surface receptor description 30-43 Nuclear receptor description 44-51 Recent advances 52,53 References 54,55 2

3. INTRODUCTION TO HORMONE A  H ormone is any member of a class of signaling molecules produced by glands in multicellular organisms that are transported by the circulatory system to target distant organs to regulate physiology and behaviour . Hormones effects on the body : S timulation or inhibition of growth Sexual arousal I nduction or suppression of  apoptosis and to control puberty and menopause A ctivation or inhibition of the immune system 3 Norman, Anthony W., and Gerald Litwack .  Hormones . Academic Press, 1997.

4. Regulation of  metabolism Control of the reproductive cycle Hormones exert their actions as: 1.Endocrine action 2.Paracrine action 3.Autocrine action INTRODUCTION TO HORMONE 4 Norman, Anthony W., and Gerald Litwack .  Hormones . Academic Press, 1997.

5. Hormone receptors have two mandatory characteristics: The receptor must identify a unique binding site within the hormone. The receptor must be able to transmit the signal caused by binding with the hormone into a cellular response. Hormonal Receptors 5 Norman, Anthony W., and Gerald Litwack .  Hormones . Academic Press, 1997.

6. Classification of Hormonal receptors 6 Giguère , Vincent, et al. "Identification of a new class of steroid hormone receptors." (1988): 91-94.

7. Structure of Cell Surface Receptors: Cell surface receptors are integral membrane proteins, have regions that contribute to three basic domains: Extracellular domains: Transmembrane domains: Cytoplasmic or intracellular domains: Cell surface receptors 7 Deller, Marc C., and E. Yvonne Jones. "Cell surface receptors."  Current opinion in structural biology  10.2 (2000): 213-219.

8. Cell surface receptors hormones can show theirs action via 2 different pathways by activating various sec. messengers systems which are, 1) G-Protein couple receptor(GPCR ) pathway. cAMP sec.messenger calcium or phosphoinositide sec.messenger 2) Tyrosine kinase receptor pathway. Mechanisms of cell surface receptors 8 Deller, Marc C., and E. Yvonne Jones. "Cell surface receptors."  Current opinion in structural biology  10.2 (2000): 213-219.

9. 1)GPCR pathway 1. cAMP sec.messenger activation 9

10. Epinephrine and Norepinephrine Glucagon Luteinizing hormone, Follicle stimulating hormone Thyroid-stimulating hormone,Calcitonin,Parathyroid hormone Antidiuretic hormone Examples of hormones act through such pathway 10

11. 1) GPCR pathway 2.calcium or phosphoinositide sec.messenger 11

12. Epinephrine and norepinephrine Angiotensin II Antidiuretic hormone Gonadotropin releasing hormone Thyroid-releasing hormone Examples of hormones act through such pathway 12

13. Protein Tyrosine kinase Protein tyrosine kinases (PTKs) are a large family of enzymes that play critical roles in signal transduction. Through transferring γ-phosphate group from ATP to protein substrates, PTKs regulate functions of these substrates. Dimerization A   dimer  is a macromolecular complex formed by two, usually non-covalently bound, macromolecules such as proteins  or nucleic acids . It is a quaternary structure of a protein. When two monomers get attached to each chemically and form dimer then its called dimerization. 2.tyrosine kinase pathway 13 Schlessinger , Joseph. "Cell signaling by receptor tyrosine kinases."  Cell  103.2 (2000): 211-225.

14. 2.tyrosine kinase pathway 14

15. Dimerization activates the kinase activity and leads to autophosphorylation , creating sites for interactions with specific effectors. Ras A key component of the RTK cascade is Ras Ras is small, monomeric G protein with very low GTPase activity. Grb2 SH2 protein that binds to phosphorylated RTKs It has three domains, one binds the RTK, the other Binds Sos Sos It is a Ras -GEF. When recruited to the membrane it activates Raf Raf Ras -GTP recruits Raf , which becomes activated as a protein kinase and initiates the MAP kinase cascade 15

16. Insulin Growth hormone Prolactin Oxytocin Erythropoietin Several growth factors Examples of hormones act through such pathway 16

17.   Nuclear receptors are intracellular proteins expressed in the nucleus of a cell . Nuclear receptors  are a family of ligand-regulated transcription factors that are activated by steroid hormones, such as estrogen and progesterone, and various other lipid-soluble signals, including retinoic acid , and thyroid hormone. Nuclear receptors (NRs) are proteins that share considerable amino acid sequence similarity in two highly conserved domains. DNA binding (DBD) and the ligand binding domains (LBD ). Nuclear receptors 17 Freedman, Leonard P. "Increasing the complexity of coactivation in nuclear receptor signaling ."  Cell  97.1 (1999): 5-8.

18. Nuclear receptors Nuclear receptor super family Class 1 steroid receptor family 1.Estrogen receptor 2.Androgen receptor 3.Progesteron receptor 4.Minerelocortocoid receptor 5.Glucocorticoid receptor 1.Thyroid receptor 2.Retinoicacid receptor 3.VitamineD receptor 4.Peroxisome proliferator activated receptor Class 2 receptor family Class 3 receptor family Orphans type of receptors 1.Farnesoicx receptor 2.Liver x receptor 18 Mangelsdorf , David J., et al. "The nuclear receptor superfamily: the second decade."  Cell  83.6 (1995): 835-839.

19. Basic components related to nuclear receptor: Zinc fingers : Zinc fingers have one or more zinc atoms gripped by a combination of four amino acids, either cysteine or histidine. a protein can grab a zinc ion and fold tightly around it. a short chain of 20-30 amino acids is enough to create a solid, stable structure. Many zinc fingers play essential roles in DNA recognition and essential role in the development of blood cells. Nuclear receptors 19 Nolte, Robert T., et al. "Differing roles for zinc fingers in DNA recognition: structure of a six-finger transcription factor IIIA complex."  Proceedings of the National Academy of Sciences  95.6 (1998): 2938-2943.

20. Heat shock proteins  ( HSP ): Are a family of proteins that are produced by cells in response to exposure to stressful conditions  such as infection ,  inflammation, exercise , exposure of the cell to toxins ( ethanol   arsenic ,  tracemetals and  ultraviolet  light),  starvation,  hypoxia. Heat-shock proteins are named according to their molecular weight. For example, Hsp60, Hsp70 and Hsp90  (Chaperones ).   Heat Shock Proteins bind to denatured proteins to prevent aggregation. Nuclear receptors 20 Freedman, Leonard P.  Molecular biology of steroid and nuclear hormone receptors . Springer Science & Business Media, 1998.

21. Coactivator and C orepressor : Transcription coregulators that activate gene transcription are referred to as  coactivators  while those that repress are known as corepressors . The mechanism of action of transcription coregulators is to modify chromatin structure and thereby make the associated DNA more or less accessible to transcription . Nuclear receptors 21 Petkovi c , Martin, et al. "A human retinoic acid receptor which belongs to the family of nuclear receptors."  Nature  330.6147 (1987): 444-450

22. A  hormone response element (HRE) is a short sequence of DNA within the promoter of a gene that is able to bind a specific hormone receptor complex and therefore regulate transcription. HRE responds to steroid hormones, as the activated steroid receptor is the transcription factor binding HRE . Nuclear receptors sequence of DNA Actual mechanism 22 Umesono , Kazuhiko, et al. "Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D 3 receptors."  Cell  65.7 (1991): 1255-1266.

23. Nuclear receptors A/B region C region D region E region F region 23 Moras , Dino, and Hinrich Gronemeyer . "The nuclear receptor ligand-binding domain: structure and function."  Current opinion in cell biology  10.3 (1998): 384-391.

24. Nuclear receptors   DNA-binding domain ( DBD ): Highly conserved domain containing two zinc fingers that binds to specific sequences of DNA called hormone response elements (HRE). Zinc atoms D ouble strained DNA DNA-binding domain dimer 24

25. Nuclear receptors Ligand binding domain ( LBD ): The structure of the LBD is referred to as an alpha helical sandwich fold in which three anti parallel alpha helices are flanked by two alpha helices on one side and three on the other. The ligand binding cavity is within the interior of the LBD and just below three anti parallel alpha helical sandwich " filling“. Crystallographic structure of the ligand binding domain of the human RORγ (rainbow colored , N-terminus = blue, C-terminus = red) complexed with 25-hydroxycholesterol (space-filling model (carbon = white, oxygen = red) and the NCOA2 coactivator ( magneta ). 25

26. Mechanisms of nuclear receptors 1) Mechanism of class I nuclear receptor action 2 6

27. Steroid hormone receptors: Glucocorticoid Receptor Mineralocorticoid Receptor Androgen Receptor Estrogen Receptor Progesterone Receptor Examples of hormones act through such mechanism 2 7

28. Mechanisms of nuclear receptors 2) Mechanism of class II nuclear receptor action 2 8

29. Retinoic acid receptor Retinoid X receptor Thyroid hormone receptor Examples of hormones act through such mechanism 2 9

30. Steroid hormone receptors : Glucocorticoid Receptor Androgen Receptor Estrogen Receptor Progesterone Receptor Thyroid hormone receptor: nuclear receptors description 3

31. History of glucocorticoid receptor: Glucocorticoid receptor (GR) drug discovery was initiated by adrenal gland extracts. In 1948 sufficient quantities of cortisone could be purified and its effects in inflammatory disease could be tested. Finally, total syntheses of cortisone were independently achieved by R.B.Woodward. Glucocorticoid Receptor 3 1 Rousseau, Guy G., John D. Baxter, and Gordon M. Tomkins. "Glucocorticoid receptors: relations between steroid binding and biological effects."  Journal of molecular biology  67.1 (1972): 99-115.

32. Agonist Antagonist Cortisol Mifepristone Corticosterone Org 34850 Aldosterone RU-486 Glucocorticoid Receptor The GR is expressed in a wide variety of lymphoid cells, including T and B cells, and macrophages. 32

33. Role of Glucocorticoid Receptor In Disease Condition : Inflammatory and immune disease Asthma therapy Allergic Reaction Myasthenia Gravis Infective Disease Hypercalcaemia Glucocorticoid Receptor 3 3

34. Androgens act via the AR and play an important role in the development and differentiation of the male sexual organ . It is also important in maintaining skeletal muscle mass and strength, BMD, hematopoiesis . The two natural occurring androgens are testosterone (T) and the more potent 5α- dihydrotestosterone (DHT). Androgen Receptor 3 4 Drews , Jürgen. "Drug discovery: a historical perspective."  Science  287.5460 (2000): 1960-1964.

35. Role of Androgen Receptor In Disease Condition Hypogonadism Anaemia Osteoporosis Androgen Receptor Agonist Antagonist  Synthetic progestin Flutamide Dihydrotestosterone Spironolactone Testosterone Danazol 3 5

36. History of estrogen receptor: Estrone was isolated in 1929 as the first natural estrogenic hormone independently by the chemists E. A. Doisy and A. Butenand from the urine of pregnant women In 1938, H. H. Inhoffen achieved for the first time the synthesis of estradiol from cholesterol. The first synthesis of ethinyl-estradiol,which has an improved oral bioavailability , was published in 1938 by H. H. Inhoffen and W . Hohlweg . Estrogen Receptor 3 6 Drews , Jürgen. "Drug discovery: a historical perspective."  Science  287.5460 (2000): 191964 .

37. Both ERs are widely expressed, although ER α is more abundant in breast , endometrial , ovarian, and hypothalamus tissues , whereas ER β is more abundant in brain , bone, endothelial, heart, intestine , kidney, lung, and prostate tissues . Estrogen Receptor Agonist Antagonist Estradiol Tamoxifen Estrone Doloxifen Estriol Toremifen Clomiphene Raloxifene 3 7

38. 3 8

39. History of progesterone receptor: The observation that progesterone prevented ovulation was taken up by Haberlandt who was able to demonstrate that it could be used for fertility control. In 1955 , Gregory Pincus demonstrated that large doses of orally Administered progesterone inhibited ovulation in women. Inhoffen had discovered in 1938 that 17a-ethynyl-testosterone was a weakly active progestin when administered orally. Progesterone Receptor 3 9 Drews , Jürgen. "Drug discovery: a historical perspective."  Science  287.5460 (2000): 1960-1964 .

40. The PR is involved in diverse functions in female reproduction Progesterone is mainly produced in the corpus luteum, placenta The PR is expressed in the uterus, ovary. Role of Progesterone Receptor In Disease Condition: Contraception Hormone Replacement Therapy Breast Cancer Endometrial Hyperplasia Progesterone Receptor 4

41. The thyroid hormones (THs) triiodothyronine (T3) and thyroxine (T4) bind to and activate the nuclear thyroid receptors TR α and TR β . Studies dating back to the 19th century showed that transplantation of sheep thyroid gland tissue could rapidly improve the state of a patient suffering from hypothyroidism. Differential binding activity of TH α and TH β could be shown among various tissues, with high binding in the liver, kidney, pituitary gland, heart, and brain. Thyroid Hormone ReceptOR 4 1 Shibata, Hirotaka , et al. "Role of co-activators and co-repressors in the mechanism of steroid/thyroid receptor action."  Recent Progress in Hormone Research  52 (1996): 141-64

42. Actions of Thyroid Hormone Receptor Growth and Development Calorigenic Effects Cardiovascular Effects Metabolic Effects Thyroid Hypofunction Thyroid Hyperfunction Thyroid Hormone ReceptOR 4 2

43. Disorders of the thyroid gland Hyperthyroidism—excess of thyroid hormone production. Hypothyroidism—deficiency of thyroid Hormone production . Goitre formation . Adenoma (benign growths) of the thyroid . Carcinoma of the thyroid . Thyroid Hormone ReceptOR 4 3 Shibata, Hirotaka , et al. "Role of co-activators and co-repressors in the mechanism of steroid/thyroid receptor action."  Recent Progress in Hormone Research  52 (1996): 141-64.

44. Growth hormone receptor Insulin receptor V asopressin receptor Cell surface receptors description 4 4

45. Growth hormone was identified in the 1920s as a growth promoting Factor. By mid to late 1980s scientists were able to produce this 191-residue protein hormone in bacteria using recombinant DNA technology. Growth hormone is a protein hormone of about 190 amino acids that is synthesized and secreted by cells called somatotrophs in the anterior pituitary . Growth hormone receptor 4 5

46. Growth hormone receptor When two receptors are brought together, interaction between the portions inside the cell triggers several enzymatic reactions and signaling processes that stimulate growth . Thus formation of this receptor dimer is crucial for growth hormone function. 4 6

47. To regulate normal growth and development of body. Metabolic Effects Protein metabolism : Fat metabolism : Carbohydrate metabolism : Role of Growth hormone in Disease States . Gigantism Acromegaly Growth hormone receptor 4 7

48. Insulin receptor I nsulin receptor visualised by electron microscope in frozen hydrated state(top) and its illustration(bottom) Structure of Insulin receptor 4 8

49. Insulin receptor 49

50. Vasopressin receptor Distribution and signaling pathway of vasopressin receptor. RECEPTOR SIGNALING PATHWAY LOCATION V 1( V 1 A) G protein couple/phosphatidylinositol calcium Vascular smooth muscle, platelet, Hepatocytes and myometrium. V 2 Adenylyl cyclase/ cAMP Basolateral membrane of collecting duct and vascular smooth muscle cell. V 3 (V 1 B) G protein couple/phosphatidylinositol calcium Anterior pituitary gland. 5 Jagadeesh , J. S., Muthiah , N. S., & Muniappan , M. (2014). Vasopressin Receptors and Drugs: A Brief Perspective.  Global Journal of Pharmacology , 8 (1), 80-83.

51. Functions of vasopressin receptor Vasoconstriction Myocardial hypertrophy Platelet aggregation Glycogenolysis Uterine contraction Insertion of AQP-2 water channels into apical membrane Releases ACTH, prolactin, endorphins Vasopressin receptor 5 1

52. 1)The concentration of the estrogen binding protein in the rat uterus has been studied in three stages of uterine development. A 3- to 4-fold increase in concentration of estrogen binding protein occurs between days one and ten after birth . After estrogen is injected, the receptor concentration in the cytoplasm goes into a three-stage cycle. (1) Initially , there is a loss of binding protein, followed by (2) a stage sensitive to inhibitors of protein and RNA synthesis, and (3) a replenishment period that is not affected by the inhibitors Recent Advances 52 Gorski, J., M. Sarff , and J. Clark. "The regulation of uterine concentration of estrogen binding protein." (1971).

53. 2)Bisphosphonates (BPs) have become the standard therapy for breast cancer patients with bone metastasis. It is now clear that the bisphosphonates reduce both the symptoms and complications of bone involvement The administered bisphosphonates (BPs) differed according to the periods of treatment, compliance, tolerability and insurance strategy. The types of bisphosphonate used were zoledronate or pamidronate . These drugs were given at three or four week intervals. Recent Advances 5 3 Park, In H., et al. "Potential antitumor effects of nitrogen-containing bisphosphonate in hormone receptor negative breast cancer patients with bone metastases."  BMC cancer  9.1 (2009): 154.

54. Norman, Anthony W., and Gerald Litwack .  Hormones . Academic Press, 1997 Giguère , Vincent, et al. "Identification of a new class of steroid hormone receptors." (1988): 91-94 . Deller, Marc C., and E. Yvonne Jones. "Cell surface receptors."  Current opinion in structural biology  10.2 (2000): 213-219 . Schlessinger , Joseph. "Cell signaling by receptor tyrosine kinases."  Cell  103.2 (2000): 211-225. Freedman, Leonard P. "Increasing the complexity of coactivation in nuclear receptor signaling ."  Cell  97.1 (1999): 5-8. Mangelsdorf , David J., et al. "The nuclear receptor superfamily: the second decade."  Cell  83.6 (1995): 835-839 . Nolte, Robert T., et al. "Differing roles for zinc fingers in DNA recognition: structure of a six-finger transcription factor IIIA complex."  Proceedings of the National Academy of Sciences  95.6 (1998): 2938-2943 . Petkovic, Martin, et al. "A human retinoic acid receptor which belongs to the family of nuclear receptors."  Nature  330.6147 (1987): 444-450 Umesono , Kazuhiko, et al. "Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D 3 receptors."  Cell  65.7 (1991): 1255-1266. Moras , Dino, and Hinrich Gronemeyer . "The nuclear receptor ligand-binding domain: structure and function."  Current opinion in cell biology  10.3 (1998): 384-391. Rousseau, Guy G., John D. Baxter, and Gordon M. Tomkins. "Glucocorticoid receptors: relations between steroid binding and biological effects."  Journal of molecular biology  67.1 Drews , Jürgen. "Drug discovery: a historical perspective."  Science  287.5460 (2000): 1960-1964. Shibata, Hirotaka , et al. "Role of co-activators and co-repressors in the mechanism of steroid/thyroid receptor action."  Recent Progress in Hormone Research  52 (1996): 141-64 refrences 54

55. Jagadeesh , J. S., Muthiah , N. S., & Muniappan , M. (2014). Vasopressin Receptors and Drugs: A Brief Perspective.  Global Journal of Pharmacology , 8 (1), 80-83 Gorski, J., M. Sarff , and J. Clark. "The regulation of uterine concentration of estrogen binding protein." (1971 ). Park, In H., et al. "Potential antitumor effects of nitrogen-containing bisphosphonate in hormone receptor negative breast cancer patients with bone metastases."  BMC cancer  9.1 (2009): 154. refrences 55