STRUCTURE/FUNCTION OF HEME REGULATED ELF-2ALPHA KINASE

血红素调节的 ELF-2α 激酶的结构/功能

• 批准号: 6350684
• 负责人: JANE-JANE CHEN
• 金额: $ 21.56万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-02-15 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6350684
• 关键词: X ray crystallography; alkylation; cell differentiation; cell growth regulation; crosslink; enzyme activity; enzyme inhibitors; enzyme structure; erythroid stem cell; erythropoiesis; heme; hemoglobin; hemoprotein biosynthesis; intermolecular interaction; mass spectrometry; molecular site; phosphorylation; protein binding; protein kinase; protein sequence; protein structure function; proteolysis; reticulocytes; site directed mutagenesis; stress proteins

DESCRIPTION: The long-term objectives are the elucidation of the regulation of hemoglobin synthesis and erythropoiesis in normal and abnormal hematologic conditions. The focus of this research is to study the mechanisms of regulation of heme-regulated EIF-2a kinase (HRI) at the molecular structural level. HRI is a potent inhibitor of protein synthesis in immature erythroid cells. Protein synthesis in intact reticulocytes and their lysates is dependent on the availability of heme. In heme-deficiency, protein synthesis is inhibited at the level of initiation due to the activation of HRI. HRI is a heme-regulated protein kinase that phosphorylates the a-subunit of EIF-2 impairs its recycling in translational initiation and results in the cessation of protein synthesis. HRI is mainly expressed in erythroid cells and the physiological role of HRI is to maintain the balanced synthesis of heme and globin chains. The specific aims of this proposed study are (1) Regulation of HRI by phosphorylation; (2) Molecular mechanisms in the regulation of HRI by heme, the prosthetic group of hemoglobin which is the principal proteins in reticulocytes; ( 3) Determination of HRI structure by X-ray crystallography. The methods to be employed are recombinant DNA techniques, polymerase chain reaction, synthesis of oligodeoxynucleotides, high pressure liquid chromatography, amino acid microsequencing, in vitro transcription and translation, protein kinase assays, gel electrophoresis, deoxynucleotide sequencing, expression of HRI and other proteins using recombinant baculovirus, immunoaffinity column chromatography, immunoprecipitation, Western blot analysis, and X-ray crystallography. HRI is a key regulator of protein synthesis, a process which is essential for the regulation of cell growth, differentiation, antiviral response and hormonal responses. Structure-function analysis of HRI will advance our knowledge not only on the regulation of protein kinases which are vital in the regulation of various cellular processes, but also on the regulation of heme proteins.

描述：长期目标是法规的阐明

项目成果

Phosphorylation of eukaryotic initiation factor 2 by heme-regulated inhibitor kinase-related protein kinases in Schizosaccharomyces pombe is important for fesistance to environmental stresses.

粟酒裂殖酵母中血红素调节抑制剂激酶相关蛋白激酶对真核起始因子 2 的磷酸化对于抵抗环境胁迫非常重要。

• DOI: 10.1128/mcb.22.20.7134-7146.2002
• 发表时间: 2002
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Zhan,Ke;Vattem,KrishnaM;Bauer,BettinaN;Dever,ThomasE;Chen,Jane-Jane;Wek,RonaldC
• 通讯作者: Wek,RonaldC

Autophosphorylation of threonine 485 in the activation loop is essential for attaining eIF2alpha kinase activity of HRI.

• DOI: 10.1021/bi034005v
• 发表时间: 2003-05
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: M. Rafie-Kolpin;A. Han;Jane-Jane Chen-Jane