DEVELOPMENTAL REGULATION OF MAMMARY GLYCOPROTEINS

乳腺糖蛋白的发育调节

• 批准号: 3226501
• 负责人: INDER K VIJAY
• 金额: $ 20.23万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-02-01 至 1994-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3226501
• 关键词: alpha glucosidase; antibody; carbohydrate metabolism; cell differentiation; enzyme biosynthesis; enzyme mechanism; genetic transcription; high performance liquid chromatography; histogenesis; hormone regulation /control mechanism; laboratory mouse; laboratory rabbit; lactation; mammary gland; molecular cloning; oligosaccharides; phosphotransferases; plasmids; posttranslational modifications; prolactin; protein biosynthesis; protein purification; transposon /insertion element

The long range objective of the proposed program is to understand molecular mechanisms that regulate the biosynthesis of N-linked glycoproteins during growth and differentiation. The model to be used is the mammary gland during its ontogeny. N-linked glycoproteins constitute the largest class of glycoproteins and participate in myriads of cellular reactions and other phenomena that are fundamental to biological recognition. Alterations in glycoprotein metabolism are associated with a variety of pathological and grave consequences for the host. The enzyme UDP-GlcNAc:Dol-P GlcNAc-l-P-transferase initiates the assembly of the oligosaccharide precursor Glc3Man9GlcNAc2-PP-dolichol for the biosynthesis of the carbohydrate component of these proteins. Following a transfer en bloc of the oligosaccharide to the nascent polypeptide, the enzymes glucosidase I and II trigger the processing of these proteins. Additional processing and post-translational modifications give rise to different subclasses, viz., high mannose, hybrid and complex glycoproteins. Throughout the reproductive life of the mammalian female, the mammary gland goes through highly ordered, cyclic changes under the influence of a variety of hormones that control its growth, differentiation, and regression. Our preliminary studies have shown that several of the key enzymes for the biosynthesis of N-linked glycoproteins in this tissue are developmentally controlled and prolactin appears to participate in this regulation. The enzymes GlcNAc-l-P-transferase and Glucosidases I and II being the early enzymes in biosynthesis and processing, appear to be excellent candidates for the regulation of N- linked glycoproteins in the mammary gland. We have purified these enzymes from the bovine gland; monospecific polyclonal antibodies raised against these enzymes show excellent tissue and species crossreactivity. In the proposed program, the genes for these enzymes will be cloned in the expression vector lambda gtll. These would then be used to select clones from a mouse mammary library in lambda gt10. This will be followed by a study of regulation of these enzymes, both in vivo and in vitro during gland development. Finally, using the antibodies, the biosynthesis and turnover of these enzymes would be investigated.

拟议程序的远距离目标是了解 调节N连接的生物合成的分子机制 生长和分化过程中的糖蛋白。 要使用的模型是 乳腺在其个体发育过程中。 N连接的糖蛋白构成 最大的糖蛋白类，参与无数细胞 反应和其他对生物学基础的现象 认出。 糖蛋白代谢的改变与 宿主的各种病理和严重后果。 这 酶UDP-GLCNAC：DOL-P GlcNAC-L-P-P-转移酶启动组装 寡糖前体GLC3MAN9GLCNAC2-PP-DOLICHOL用于 这些蛋白质的碳水化合物成分的生物合成。 下列的 将寡糖转移到新生多肽的转移 酶I和II酶I和II触发这些蛋白质的加工。 其他处理和翻译后修饰会引起 不同的子类别，即高甘露糖，混合和复杂 糖蛋白。 在哺乳动物女性的生殖生活中 乳腺经过高度有序的，循环的变化 控制其生长的多种激素的影响， 分化和回归。 我们的初步研究表明 N连接糖蛋白的生物合成的几种关键酶 在该组织中，受发育控制，催乳素似乎 参与此法规。 酶GlcNAC-L-P-P-转移酶和 葡萄糖酶I和II是生物合成中的早期酶 处理，似乎是调节n-的出色候选者 乳腺中连接的糖蛋白。 我们已经净化了这些 来自牛腺的酶；单特异性多克隆抗体升高 针对这些酶显示出极好的组织和物种交叉反应性。 在拟议的程序中，这些酶的基因将被克隆 表达矢量lambda gtll。 然后这些将用于选择 来自Lambda GT10的小鼠乳房库的克隆。 这将是 然后研究对这些酶的调节，包括体内和中 腺体发育过程中的体外。 最后，使用抗体 将研究这些酶的生物合成和周转。