CARBOHYDRATE BINDING PROTEIN 35

碳水化合物结合蛋白 35

• 批准号: 3295375
• 负责人: JOHN L WANG
• 金额: $ 9.58万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3295375
• 关键词: N acetylglucosamine; affinity chromatography; binding proteins; cell nucleus; complementary DNA; cytoplasm; galactose; genetic manipulation; genetic transcription; heterogeneous nuclear ribonucleoprotein; laboratory mouse; laboratory rabbit; laboratory rat; messenger RNA; molecular cloning; nuclear runoff assay; phosphoproteins; protein sequence; radiotracer; ribonucleoproteins

This project propose to study the structure and activities of Carbohydrate-Binding Protein 35 (Mr 35,000), a lectin that binds specifically to galactose-N-acetylglucosamine (Gal-GlcNAc) containing glycoconjugates. Using a highly specific antibody directed against CBP35, we have made the striking observation that CBP35 can be found in the nucleus of a cell and that the level of expression of CBP35 and its nuclear localization may be dependent on the proliferation state of the cell. More recently, we have accumulated evidence to indicate that CBP35 is a protein associated with the core particle of the heterogeneous nuclear ribonucleoprotein complex (hnRNP). The specific objectives of the proposed research are: (a) to identify, isolate, and characterize cDNA and genomic clones for CBP35; (b) to determine the partial amino acid sequence of CBP35 at the protein level and to derive the complete amino acid sequence from the cDNA sequence; (c) to study the regulation of transcription of the CBP35 gene by Northern analysis and nuclear run-on experiments; (d) to analyze CBP35 in terms of the possibility of different nuclear versus cytoplasmic forms, their relationships to the different isoelectric variants of CBP35 and the different states of phosphorylation; (e) to search for conditions that will release CBP35 from the hnRNP core particle and to identify the correspondence between CBP35 and the protein(s) of the core particle; and (f) to analyze the effect of CBP35 in certain perturbation experiments (e.g. addition of CBP35 and/or rabbit anti-CBP35) on cell free mRNA processing (splicing) systems. All of these studies should contribute to our understanding of the structure and activities of lectins endogenous to animal cells.

本项目拟研究 碳水化合物结合蛋白35（Mr 35，000），一种结合 特异性针对半乳糖-N-乙酰葡糖胺（Gal-GlcNAc） 含有糖缀合物。 使用高度特异性的抗体 针对CBP 35，我们做出了惊人的观察， CBP 35可以在细胞核中找到， CBP 35的表达水平及其核定位可能是 这取决于细胞的增殖状态。 最近， 我们积累的证据表明，CBP 35是一种蛋白质， 与异质核的核心粒子有关 核糖核蛋白复合物（hnRNP）。 拟议研究的具体目标是： (a)鉴定、分离和表征cDNA和基因组 CBP 35的克隆; (b)为了确定CBP 35的部分氨基酸序列， 蛋白质水平，并推导出完整的氨基酸序列 从cDNA序列; (c)研究CBP 35基因转录的调控， 北方分析和核连续试验 (d)分析CBP 35在不同的可能性， 细胞核与细胞质的形式，它们与 CBP 35的不同等电点变体和 磷酸化; (e)搜索将CBP 35从 hnRNP核心颗粒，并确定之间的对应关系 CBP 35和核心颗粒的蛋白质;以及 (f)分析了在一定的扰动下CBP 35的作用 实验（例如添加CBP 35和/或兔抗CBP 35） 无细胞mRNA加工（剪接）系统。 所有这些研究都有助于我们理解 动物细胞内源凝集素的结构和活性。