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Molecular cloning and expression of the pituitary glycoprotein hormone N-acetylgalactosamine-4-O-sulfotransferase

JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第49期275卷 38402-38409页

作者： Xia, GQ Evers, MR Kang, HG Schachner, M Baenziger, JU Washington Univ Sch Med Dept Pathol St Louis MO 63110 USA Univ Hamburg Zentrum Mol Neurobiol D-20246 Hamburg Germany

N-Linked oligosaccharides terminating with the sequence SO4-4-GalNAc beta1,4GlcNAc beta1,2Man alpha are present on the pituitary hormones lutropin (LH), thyrotropin, and pro-opiomelanocortin. The sulfated structures on LII are essential for expression of its biologic function in vivo. We have cloned the N-acetglgalactosamine-4-sulfo-transferase (GalNac-4-ST1, GenBank(TM) accession number AF300612), which mediates sulfate addition to the N-linked oligosaccharides on LH and other pituitary glycoproteins with terminal (beta1,4-linked GalNAc based on its homology to HNK-1 sulfotransferase (HNK-1 ST). GalNAc-4-ST1 displays 23% identity to HNK-1 ST and 28% to chondroitin 4-sulfotransferase I (C4ST-1) and 26% to chondroitin 4-sulfotransferase 2 (C4ST-2), The cDNA predicts a type II transmembrane protein of 424 amino acids with four potential N-linked glycosylation sites and a single membrane-spanning domain. GalNAc-4-ST1 has putative 5'-phosphosulfonate and S'-phosphate binding sites, Three more carboxyl-terminal regions of unknown function also show a high degree of identity with HNK-1 ST, C4ST-1, and C4ST-2. The membrane-bound form of GalNAc-4-ST1 transfers sulfate to GalNAc beta1,4GlcNAc beta -R but not to chondroitin, whereas truncated forms of GalNAc-4-ST1 that are released into the medium transfer sulfate to both GalNAc beta1, 4GlcNAc beta -R and chondroitin, The first 118 amino acids of GalNAc-4-ST1 appear to contribute to both its activity and specificity for terminal beta1,4-linked GalNAc. GalNAc-4-ST1 also efficiently transfers sulfate to N-linked oligosaccharides on native LH and other glycoproteins terminating with beta1,4-linked GalNAc. A single transcript of 2.4 kilobases is most highly expressed in the pituitary and other regions of the central nervous system. The GalNAc-4-ST1 gene is located on human chromosome 19q13.1.

关键词：氨基酸序列碱基序列 CHO细胞碳水化合物序列染色体图染色体人 19对克隆分子仓鼠亚科促黄体激素/代谢分子序列数据寡糖类/化学垂体激素类前叶/化学垂体激素类前叶/代谢前阿片黑素细胞皮质激素/代谢重组蛋白质类/代谢序列比对序列同源性氨基酸磺基转移酶类/遗传学磺基转移酶类/代谢促甲状腺素/代谢转染动物人类大鼠

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Corticotropin releasing hormone and proopiomelanocortin involvement in the cutaneous response to stress

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PHYSIOLOGICAL REVIEWS 2000年第3期80卷 979-1020页

作者： Slominski, A Wortsman, J Luger, T Paus, R Solomon, S Loyola Univ Med Ctr Dept Pathol Maywood IL 60153 USA So Illinois Univ Dept Med Springfield IL USA Univ Munster Ludwig Boltzmann Inst Cell Biol & Immunobiol Skin Dept Dermatol D-4400 Munster Germany Univ Hamburg Hosp Eppendorf Dept Dermatol D-20246 Hamburg Germany McGill Univ Dept Med & Biochem Montreal PQ Canada Royal Victoria Hosp Montreal PQ H3A 1A1 Canada

The skin is a known target organ for the proopiomelanocortin (POMC)-derived neuropeptides alpha-melanocyte stimulating hormone (alpha-MSH), beta-endorphin, and ACTH and also a source of these peptides. Skin expression levels of the POMC gene and POMC/corticotropin releasing hormone (CRH) peptides are not static but are determined by such factors as the physiological changes associated with hair cycle (highest in anagen phase), ultraviolet radiation (UVR) exposure, immune cytokine release, or the presence of cutaneous pathology. Among the cytokines, the proinflammatory interleukin-1 produces important upregulation of cutaneous levels of POMC mRNA, POMC peptides, and MSH receptors;UVR also stimulates expression of all the components of the CRH/POMC system including expression of the corresponding receptors. Molecular characterization of the cutaneous POMC gene shows mRNA forms similar to those found in the pituitary, which are expressed together with shorter variants. The receptors for POMC peptides expressed in the skin are functional and include MC1, MC5 and mu-opiate, although most predominant are those of the MC1 class recognizing MSH and ACTH. Receptors for CRH are also present in the skin. Because expression of, for example, the MC1 receptor is stimulated in a similar dose-dependent manner by UVR, cytokines, MSH peptides or melanin precursors, actions of the ligand peptides represent a stochastic (predictable) nonspecific response to environmental/endogenous stresses. The powerful effects of POMC peptides and probably CRH on the skin pigmentary, immune, and adnexal systems are consistent with stress-neutralizing activity addressed at maintaining skin integrity to restrict disruptions of internal homeostasis. Hence, cutaneous expression of the CRH/POMC system is highly organized, encoding mediators and receptors similar to the hypothalamic-pituitary-adrenal (HPA) axis. This CRH/POMC skin system appears to generate a function analogous to the HPA axis, that in the

关键词：促肾上腺皮质素释放激素/代谢毛囊/代谢前阿片黑素细胞皮质激素/代谢受体促肾上腺皮质激素/代谢受体促肾上腺皮质素释放激素/代谢受体黑皮质素信号传导/生理学皮肤/代谢应激生理学/代谢动物人类

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Specific protein-protein interaction between basic helix-loop-helix transcription factors and homeoproteins of the Pitx family

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MOLECULAR AND CELLULAR BIOLOGY 2000年第13期20卷 4826-4837页

作者： Poulin, G Lebel, M Chamberland, M Paradis, FW Drouin, J Inst Rech Clin Montreal Genet Mol Lab Montreal PQ H2W 1R7 Canada

Homeoproteins and basic helix-loop-helix (bHLH) transcription factors are known for their critical role in development and cellular differentiation. The pituitary pro-opiomelanocortin (POMC) gene is a target for factors of both families. Indeed, pituitary-specific transcription of POMC depends on the action of the homeodomain-containing transcription factor Pitx1 and of bHLH heterodimers containing NeuroD1. We now show lineage-restricted expression of NeuroD1 in pituitary corticotroph cells and a direct physical interaction between bHLH heterodimers and Pitx1 that results in transcriptional synergism. The interaction between the bHLH and homeodomains is restricted to ubiquitous (class A) bHLH and to the Pitx subfamily. Since bHLH heterodimers interact with Pitx factors through their ubiquitous moiety, this mechanism may be implicated in other developmental processes involving bHLH factors, such as neurogenesis and myogenesis.

关键词：氨基酸序列碱性螺旋-环-螺旋转录因子类结合部位 DNA结合蛋白质类/遗传学 DNA结合蛋白质类/代谢基因表达调控发育期螺旋-环-螺旋构型同源盒结构域蛋白质类/遗传学同源盒结构域蛋白质类/代谢小鼠转基因分子序列数据突变成对盒转录因子类垂体/细胞学垂体/生长和发育垂体/代谢前阿片黑素细胞皮质激素/遗传学前阿片黑素细胞皮质激素/代谢 TCF转录因子类反式激活因子类/遗传学反式激活因子类/代谢转录因子7样1蛋白转录因子/遗传学转录因子/代谢转录遗传动物小鼠

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NEW MEDICATIONS FOR DRUG ABUSE 2000年第1期909卷 186-216页

作者： Kreek, MJ Rockefeller Univ Lab Biol Addict Dis New York NY 10021 USA

In 1963, Professor Vincent P, Dole at the Rockefeller University formed a small team to develop a pharmacotherapy for the management of heroin addiction. They hypothesized that heroin addiction is a disease of the brain with behavioral manifestations, and not merely a personality disorder or criminal behavior and began to address the specific question of whether a long-acting opioid agonist could be used in the long-term maintenance treatment of heroin addiction. Over the next 35 years, many studies documented the safety, efficacy and effectiveness of methadone pharmacotherapy for heroin addiction, but Federal regulations and stigmatization of heroin addiction prevented implementation of treatment. Finally, in 1999, NIH published a report unequivocally supporting methadone maintenance pharmacotherapy for heroin addiction. Two other effective opioid agonist treatments have been developed: the even longer acting opioid agonist 1-alpha-acetylmethadol (LAAM) has been approved for pharmacotherapy for heroin addiction, and still under study is the opioid partial agonist-antagonist buprenorphine-naloxone combination. ii variety of studies, both laboratory based and clinical, have revealed the mechanisms of action of long-acting opioid agonists in treatment, including prevention of disruption of molecular, cellular and physiologic events and, In fact, allowing normalization of those functions disrupted by chronic heroin use. Recent molecular biological studies have revealed single nucleotide polymorphisms of the human mu opioid receptor gene;the mu opioid receptor is the site of action of heroin, the major opiate drug of abuse, analgesic agents such as morphine, and the major treatment agents for heroin addiction. These findings support the early hypotheses of our laboratory that addiction may be due to a combination of genetic, drug-induced and environmental (including behavioral) factors and also, that atypical stress responsivity may contribute to the acquisition and per

关键词：促肾上腺皮质激素/代谢促肾上腺皮质素释放激素/代谢海洛因依赖/药物疗法海洛因依赖/代谢下丘脑-垂体系统/药物作用美沙酮/药理学美沙酮/治疗应用醋美沙朵/治疗应用垂体肾上腺系统/药物作用前阿片黑素细胞皮质激素/代谢 β内啡肽/生理学人类

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Cushing's syndrome: from patients to proteins

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EUROPEAN JOURNAL OF ENDOCRINOLOGY 2000年第3期143卷 313-315页

作者： Shomali, ME Hussain, MA Harvard Univ Massachusetts Gen Hosp Sch Med Endocrine Unit Boston MA 02114 USA Harvard Univ Massachusetts Gen Hosp Sch Med Mol Endocrinol Lab Boston MA 02114 USA

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Activation and routing of membrane-tethered prohormone convertases 1 and 2

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JOURNAL OF BIOLOGICAL CHEMISTRY 1999年第35期274卷 24703-24713页

作者： Bruzzaniti, A Marx, R Mains, RE Johns Hopkins Univ Sch Med Dept Neurosci Baltimore MD 21205 USA

Many peptide hormones and neuropeptides are processed by members of the subtilisin-like family of prohormone convertases (PCs), which are either soluble or integral membrane proteins. PC1 and PC2 are soluble PCs that are primarily localized to large dense core vesicles in neurons and endocrine cells. We examined whether PC1 and PC2 were active when expressed as membrane-tethered proteins, and how tethering to membranes alters the biosynthesis, enzymatic activity, and intracellular routing of these PCs. PC1 and PC2 chimeras were constructed using the transmembrane domain and cytoplasmic domain of the amidating enzyme, peptidylglycine cu-amidating monooxygenase (PAM). The membrane-tethered PCs were rerouted from large dense core vesicles to the Golgi region. In addition, the chimeras were transiently expressed at the cell surface and rapidly internalized to the Golgi region in a fashion similar to PAM. Membrane-tethered PC1 and PC2 exhibited changes in pro domain maturation rates, N-glycosylation, and in the pH and calcium optima required for maximal enzymatic activity against a fluorogenic substrate. In addition, the PC chimeras efficiently cleaved endogenous pro-opiomelanocortin to the correct bioactive peptides. The PAM transmembrane domain/cytoplasmic domain also prevented stimulated secretion of pro-opiomelanocortin products in AtT-20 cells.

关键词：促肾上腺皮质激素/代谢天冬氨酸内肽酶类/代谢钙/代谢细胞系酶激活荧光抗体技术基因 myc 糖基化高尔基体/代谢氢离子浓度膜蛋白质类/代谢混合功能氧化酶类多酶复合物前阿片黑素细胞皮质激素/代谢前蛋白转化酶1 前蛋白转化酶2 前蛋白转化酶类重组融合蛋白质类/遗传学溶解度底物特异性枯草杆菌蛋白酶类/代谢转染动物人类小鼠

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Obesity in the mouse model of pro-opiomelanocortin deficiency responds to peripheral melanocortin

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NATURE MEDICINE 1999年第9期5卷 1066-1070页

作者： Yaswen, L Diehl, N Brennan, MB Hochgeschwender, U Oklahoma Med Res Fdn Dev Biol Program Oklahoma City OK 73104 USA NIMH Mol Genet Unit Clin Neurosci Branch Bethesda MD 20892 USA Eleanor Roosevelt Inst Canc Res Denver CO 80206 USA

Pro-opiomelanocortin (POMC)-derived peptides (the melanocortins adrenocorticotropin, alpha-, beta- and gamma-melanocyte stimulating hormone;and the endogenous opioid beta-endorphin) have a diverse array of biological activities, including roles in pigmentation, adrenocortical function and regulation of energy stores, and in the immune system and the central and peripheral nervous systems'. We show here that mice lacking the POMC-derived peptides have obesity, defective adrenal development and altered pigmentation. This phenotype is similar to that of the recently identified human POMC-deficient patients(2). When treated with a stable alpha-melanocyte-stimulating hormone agonist, mutant mice lost more than 40% of their excess weight after 2 weeks. Our results identify the POMC-null mutant mouse as a model for studying the human POMC-null syndrome, and indicate the therapeutic use of peripheral melanocortin in the treatment of obesity.

关键词：肾上腺/药物作用肾上腺/病理学肾上腺/病理生理学促肾上腺皮质激素/缺乏促肾上腺皮质激素/遗传学促肾上腺皮质激素/代谢醛固酮/血液儿茶酚胺类/血液皮质酮/血液疾病模型动物基因缺失毛发颜色瘦素肥胖症/血液肥胖症/药物疗法肥胖症/病理生理学表型前阿片黑素细胞皮质激素/缺乏前阿片黑素细胞皮质激素/遗传学前阿片黑素细胞皮质激素/代谢蛋白质类/分析体重减轻/药物作用 α促黑素/激动剂 α促黑素/类似物和衍生物 α促黑素/药理学 α促黑素/治疗应用动物女(雌)性人类小鼠

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PROOPIOMELANOCORTIN IN THE HELMINTH SCHISTOSOMA-MANSONI - SYNTHESIS OF BETA-ENDORPHIN, ACTH, AND ALPHA-MSH - EXISTENCE OF POMC-RELATED SEQUENCES

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 1992年第1期650卷 245-250页

作者： DUVAUXMIRET, O CAPRON, A CTR PAUL BROCA INSERM U159 F-75013 PARIS FRANCE

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