Determining the scope of prenylatable protein sequences

确定可异戊二烯化的蛋白质序列的范围

• 批准号: 10461733
• 负责人: Natarajan Kannan
• 金额: $ 38.67万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10461733
• 关键词: Address; Amino Acid Sequence; Amino Acids; Belief; Biochemical; Bioinformatics; Biological; Biology; CENP-E protein; Charge; Complex; Consensus; Coupled; Cysteine; Data; Disease; Enzymes; Frequencies; Genetic; Guanosine Triphosphate Phosphohydrolases; Human; In Vitro; Investigation; Knowledge; Lead; Machine Learning; Mediating; Membrane; Methods; Methylation; Modification; Molecular; Molecular Chaperones; Monitor; Nuclear Lamin; Online Systems; Partner in relationship; Pathway interactions; Phenotype; Pheromone; Phosphotransferases; Positioning Attribute; Post Translational Modification Analysis; Post-Translational Protein Processing; Prevalence; Property; Protein Farnesylation; Proteins; Proteolysis; Reporter; Reporting; Research; Shapes; Specificity; Testing; Therapeutic; Ursidae Family; Yeasts; base; biophysical analysis; farnesylation; genetic selection; in vivo; isoprenylation; next generation sequencing; prediction algorithm; prevent; protein farnesyltransferase; protein function; protein geranylgeranyltransferase; protein protein interaction; ras-Related G-Proteins

PROJECT SUMMARY This study investigates the specificity of farnesyl transferase (FTase) that isoprenylates CaaX proteins. Studies probing the in vivo activity of the FTase have historically used reporters (e.g. Ras GTPases) that undergo complex multi-step post-translational modification (PTM), involving initial farnesylation followed by CaaX proteolysis and carboxyl methylation. This study takes advantage of Hsp40 Ydj1, a farnesylation-only reporter for which we have developed a range of methods to monitor its PTM status. Its use reveals that farnesylation is not necessarily coupled to subsequent PTMs as has been generally accepted for CaaX proteins, and that FTase specificity is significantly more promiscuous than anticipated. These findings challenge the conventional paradigm for how farnesylated proteins are modified and which proteins are targeted by FTase. We will extend our studies to fully resolve the specificity of FTase using a combination of genetic, biochemical, bioinformatic, and biophysical studies. We bring to bear on our investigations an exceptionally strong set of preliminary findings, the complementary expertise of several research groups, and a comprehensive molecular toolbox for the study of farnesylated proteins and other enzymes associated with this post-translational modification pathway.

项目总结

项目成果

A comprehensive in vivo screen of yeast farnesyltransferase activity reveals broad reactivity across a majority of CXXX sequences.

• DOI: 10.1093/g3journal/jkad094
• 发表时间: 2023-07-05
• 期刊: G3 (Bethesda, Md.)

Functional classification and validation of yeast prenylation motifs using machine learning and genetic reporters.

• DOI: 10.1371/journal.pone.0270128
• 发表时间: 2022
• 期刊: PloS one
• 影响因子: 3.7

Specific Disruption of Ras2 CAAX Proteolysis Alters Its Localization and Function.

• DOI: 10.1128/spectrum.02692-22
• 发表时间: 2023-02-14
• 期刊: Microbiology spectrum
• 影响因子: 3.7