CARBOHYDRATE BINDING PROTEIN 35

本项目拟研究的是 碳水化合物结合蛋白35(Mr 35,000)，一种结合凝集素的 特别是半乳糖-N-乙酰氨基葡萄糖(Gal-GlcNAc) 含有糖共轭化合物的。使用一种高度特异的抗体 针对CBP35，我们进行了引人注目的观察 CBP35可以在细胞的细胞核中找到，而且 CBP35的表达水平及其核定位可能是 取决于细胞的增殖状态。最近， 我们已经积累的证据表明CBP35是一种蛋白质 与非均相核的核心粒子有关 核糖核蛋白复合体(HnRNP)。 拟议研究的具体目标是： (A)鉴定、分离和鉴定cDNA和基因组 CBP35克隆； (B)测定CBP35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cDNAs序列中克隆出； (C)研究CBP35基因转录调控。 北方分析和核补给实验； (D)分析CBP35是否有可能不同 核型与细胞质的关系，以及它们与 CBP35的不同等电变异体及其不同状态 磷酸化； (E)搜索将CBP35从 HnRNP核心粒子与识别之间的对应关系 CBP35和核心颗粒的蛋白质(S)； (F)分析CBP35在一定扰动下的影响 试验(例如，添加CBP35和/或兔抗CBP35) 无细胞信使核糖核酸加工(剪接)系统。 所有这些研究都应该有助于我们理解 动物细胞内源性凝集素的结构和活性。

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.