REGULATION OF PHOSPHOINOSITIDE METABOLISM & EARLY RESPONSE TO OSMOTIC STRESS

磷酸肌醇代谢的调节

• 批准号: 6258790
• 负责人: WENDY F BOSS
• 金额: $ 0.05万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6258790
• 关键词: biological products; biomedical equipment development; biomedical resource; biotechnology; proteins; technology /technique development

One goal of our project is to understand the mechanism by which elongation factor 1A (eEF-1A) regulates phosphatidylinositol 4-kinase activity. eEF-1A is a multifunctional protein. There are three known posttranslational modifications of eEF-1A which could potentially affect its function. Except for phosphorylation, the other posttranslational modifications have not been demonstrated in plants. We used MALDI-MS to show that carrot eEF-1A contains a phosphoglycerylethanolamine (PGE) posttranslational modification and to identify the peptide fragment which is modified. While we could demonstrate by in vivo labeling studies that eEF-1A incorporated [14C]ethanolamine it was only by using mass spectrometry that we could definitely characterize the postranslational modification. Our data indicate that the PGE posttranslational modification alone does not affect the function of eEF-1A; however, in combination with other posttranslational modifications, the PGE lipid modification could affect the distribution and thereby affect the function of eEF-1A within the cell. We are continuing our studies using site-directed mutagenesis to knock out the posttranslationally modified amino acid. The mass spectra data were essential for this work.

我们项目的一个目标是了解 延伸因子1A（eEF-1A）调节磷脂酰肌醇4-激酶 活动 eEF-1A是一种多功能蛋白。 已知有三个 eEF-1A的翻译后修饰， 影响其功能。 除了磷酸化， 翻译后修饰在植物中尚未得到证实。 我们使用MALDI-MS显示胡萝卜eEF-1A含有一个 磷酸甘油乙醇胺（PGE）翻译后修饰和 以鉴定被修饰的肽片段。 虽然我们可以 通过体内标记研究证明， [14 C]乙醇胺，只有通过使用质谱，我们才能 明确地描述了翻译后的修改。 我们的数据 表明PGE翻译后修饰本身并不 影响eEF-1A的功能;然而，与其他结合 翻译后修饰，PGE脂质修饰可以 影响分布，从而影响eEF-1A的功能 在细胞内。 我们正在继续我们的研究， 诱变以敲除后修饰的氨基酸。 质谱数据对这项工作至关重要。