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SUBCELLULAR LOCALIZATION OF TSH PROCESSING

SUBCELLULAR LOCALIZATION OF TSH PROCESSING
TSH 加工的亚细胞定位
批准号:
3446184
负责人:
JAMES MAGNER
金额:
$5.13万
依托单位国家:
美国
项目类别:
财政年份:
1985
资助国家:
美国
项目状态:
已结题
起止时间:
1985-07-01 至 1988-06-30

项目摘要

项目成果

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中文摘要
翻译
提出的研究目的是确定亚细胞
英文摘要
The objective of the proposed research is to determine the subcellular locations of the post-translational processing of thyroid-stimulating hormone (TSH) subunit precursors. Mouse thyrotropic tumor will be utilized as a source of actively secreting thyrotrophs; pituitary tissue of euthyroid and thyroidectomized mice will be sources of "resting" and "stimulated" thyrotrophs, respectively. Tumor and pituitary tissue minces will be incubated in vitro with [S-35] methionine, [H-3] fucose, [S-35] sulfate, [H-3]N- acetylmannosamine, or [H-3] mannose, and then homogenized and fractionated by the sucrose step-gradient method that we have used for previous studies. Cell fractions enriched in rough endoplasmic reticulum, proximal Golgi, distal Golgi, and secretory granules will be characterized by electron microscopy, and assayed for "organelle-specific" enzymes, such as galactosyltransferase. The sequential movement of labeled precursors between these compartments during pulse-chase studies will be analyzed. Carboxyl cyanide m-chlorophenylhydrazone (CCCP) and monensin will be used as adjuncts to subcellular fractionation to make inferences about the subcellular location of processing events. Labeled TSH subunits will be immunoprecipitated from cell homogenates, cell fractions, and media, and will be characterized by SDS-polyacrylamide gel electrophoresis. [H-3] mannose-labeled oligosaccharides will be released from TSH subunits by endoglycosidase H, and their structures determined by paper chromatography. We will determine whether the three tissue types differ in their distribution of newly synthesized TSH subunits among subcellular compartments, the locations or kinetics of processing events, or the translocation of subunits between compartments. We will determine for each compartment of each tissue type the extent and kinetics of alpha-beta subunit combination, the molar ratios of subunits, the subunit content of certain residues (fucose, sialic acid, and sulfate), and the structures of the high mannose oligosaccharides of subunits. Unlike studies of glycoprotein processing by others, we propose to study a hormonally-responsive system in which the oligosaccharides of a major secreted species, TSH, are believed to be important for biological activity. Our findings will be clinically relevant to rare patients with hypothalamic hypothyroidism and pituitary TSH-producing tumors. Primarily, we hope to contribute to a better understanding of the basic cell biology of TSH biosynthesis.
期刊论文(11)
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Rates of processing of the high mannose oligosaccharide units at the three glycosylation sites of mouse thyrotropin and the two sites of free alpha-subunits.
小鼠促甲状腺素的三个糖基化位点和游离 α 亚基的两个位点处的高甘露糖寡糖单元的加工速率。
DOI: 10.1210/endo-123-3-1296
发表时间: 1988
期刊: Endocrinology
影响因子: 4.8
作者: [Miura,Y, Perkel,VS, Magner,JA]
通讯作者: Magner,JA
Blockade by brefeldin A of intracellular transport of secretory proteins in mouse pituitary cells: effects on the biosynthesis of thyrotropin and free alpha-subunits.
布雷菲德菌素 A 阻断小鼠垂体细胞中分泌蛋白的细胞内转运:对促甲状腺素和游离 α 亚基生物合成的影响。
DOI: 10.1210/endo-122-3-912
发表时间: 1988
期刊: Endocrinology
影响因子: 4.8
作者: [Magner,JA, Papagiannes,E]
通讯作者: Papagiannes,E
Differential susceptibility to N-glycanase at the individual glycosylation sites of mouse thyrotropin and free alpha-subunits.
小鼠促甲状腺素和游离 α 亚基的各个糖基化位点对 N-聚糖酶的敏感性不同。
DOI: 10.1210/endo-123-5-2207
发表时间: 1988
期刊: Endocrinology
影响因子: 4.8
作者: [Miura,Y, Perkel,VS, Magner,JA]
通讯作者: Magner,JA
Subcellular localization of fucose incorporation into mouse thyrotropin and free alpha-subunits: studies employing subcellular fractionation and inhibitors of the intracellular translocation of proteins.
岩藻糖掺入小鼠促甲状腺素和游离 α 亚基的亚细胞定位:采用亚细胞分级和蛋白质细胞内易位抑制剂的研究。
DOI: 10.1210/endo-119-3-1315
发表时间: 1986
期刊: Endocrinology
影响因子: 4.8
作者: [Magner,JA, Novak,W, Papagiannes,E]
通讯作者: Papagiannes,E
共 9 条
    SUBCELLULAR LOCALIZATION OF TSH PROCESSING
    SUBCELLULAR LOCALIZATION OF TSH PROCESSING
    • 批准号:
      3462843
    • 项目类别:
    • 资助金额:
      $9.0万
    • 财政年份:
      1987
    • 负责人:
      JAMES MAGNER
    • 依托单位:
    SUBCELLULAR LOCALIZATION OF TSH PROCESSING
    SUBCELLULAR LOCALIZATION OF TSH PROCESSING
    海外基金