STUDIES ON THE NATURE AND FUNCTION OF THE PHOSPHOPROTEIN, PROSOLIN

磷酸蛋白普罗索林的性质和功能研究

• 批准号: 3813401
• 负责人: H L COOPER
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3813401
• 关键词: DNA replication; cell differentiation; cell growth regulation; cellular oncology; complementary DNA; cytoskeletal proteins; gel electrophoresis; genetic library; growth inhibitors; hematopoietic stem cells; human subject; human tissue; leukocyte activation /transformation; lymphocyte; molecular cloning; myelogenous leukemia; neoplastic cell; nucleic acid sequence; oligonucleotides; phorbols; phosphoproteins; phosphorylation; protein biosynthesis; protein sequence; protein structure function; tissue /cell culture

We have extensively characterized prosolin, a major cytosolic protein of proliferating HL-60 promyelocytic leukemia cells that undergoes very rapid phosphorylation in response to treatment of the cells with tumor-promoting phorbol esters like TPA as an early step in a pathway leading to cessation of cell growth and onset of monocytoid differentiation. Prosolin has been identified in proliferating human peripheral blood lymphocytes (PBL) by tryptic peptide mapping of proteins isolated from two-dimensional electrophoretic gels. While virtually absent from resting PBL, prosolin expression is induced by mitogens as a late event, during the onset of S-phase. Its level of synthesis correlates with the prevalent level of DNA synthesis. Prosolin undergoes rapid phosphorylation in proliferating PBL in response to TPA or A23187 treatment, together with an inhibition of DNA synthesis. Phosphorylation of prosolin may be part of the mechanism by which naturally occurring growth inhibitory factors terminate normal lymphocyte proliferation. T-cell leukemia cell lines show a marked reduction in the phosphorylation of prosolin in response to A23187 and other agents. We are cloning and sequencing cDNAs to prosolin. Partial amino acid sequence data were obtained by analysis of tryptic peptides of prosolin isolated from two-dimensional electrophoretic gels. Degenerate oligonucleotide probes were synthesized and used to probe a lymphoblast cDNA library.

我们已经广泛地表征了前溶胶蛋白，一种主要的细胞溶质蛋白， 增殖的HL-60早幼粒细胞白血病细胞经历非常快速的 细胞对肿瘤促进剂处理的反应中的磷酸化 佛波酯如TPA作为导致戒烟途径的早期步骤 细胞生长和单核细胞样分化的开始。Prosolin已被 在增殖的人外周血淋巴细胞（PBL）中， 胰蛋白酶肽图谱的蛋白质分离的二维 电泳凝胶。虽然在静息PBL中几乎不存在，但前溶胶蛋白 表达是由有丝分裂原诱导的晚期事件，在 S期它的合成水平与DNA的普遍水平相关 合成. Prosolin在增殖的PBL中经历快速磷酸化， 对TPA或A23187处理的响应，以及DNA的抑制 合成.前溶胶蛋白的磷酸化可能是通过以下机制的一部分： 自然发生的生长抑制因子终止正常的 淋巴细胞增殖T细胞白血病细胞系显示出明显的 响应A23187的前溶胶蛋白磷酸化的减少， 其他代理人。我们正在克隆和测序前溶胶蛋白的cDNA。部分 氨基酸序列数据是通过分析 从二维电泳凝胶中分离的前溶胶蛋白。简并 合成寡核苷酸探针并用于探测淋巴母细胞 cDNA文库