Molecular mechanisms of protein glycosylation and trafficking

蛋白质糖基化和运输的分子机制

基本信息

批准号：
10655796
负责人：
Huilin Li
金额：
$ 47.5万
依托单位：
VAN ANDEL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-18 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10655796
关键词：
ADP-Ribosylation Factors Address Back Binding Biogenesis Biological Process C-terminal Catalytic Domain Cell membrane Cells Clathrin-Coated Vesicles Complex Couples Coupling Development Docking Dolichyl-phosphate-mannose-protein mannosyltransferase Early Endosome Endosomes Enzymes Eukaryota Family Family member Funding GTPase-Activating Proteins Goals Guanine Nucleotide Exchange Factors Guanine Nucleotides Guanosine Triphosphate Phosphohydrolases Human Hydrophobicity In Vitro Knowledge Link Lipid Bilayers Lipids Location Lysosomes Mannosyltransferases Mediating Membrane Membrane Proteins Molecular Monomeric GTP-Binding Proteins N-terminal Pathologic Phosphatidylserines Phosphotransferases Play Process Protein Glycosylation Proteins Proteome Resolution Ribosomes Role Rough endoplasmic reticulum Series Signal Transduction Sorting Structure Tail Testing Therapeutic Transcription Factor AP-1 Transmembrane Domain Travel Vesicle Yeasts alpha helix cancer therapy dimer glycosylation glycosyltransferase mannose 6 phosphate member polypeptide prevent protein complex protein structure protein transport protein-O-mannosyltransferase 1 protein-O-mannosyltransferase 2 reconstitution recruit small molecule trafficking trans-Golgi Network tumor progression tumorigenesis vesicle transport

项目摘要

Project Summary The objective of this proposal is to gain mechanistic and pathological understanding of protein glycosylation and trafficking. During the previous funding cycle, we have made important contributions to this process. We have determined the structure of the transmembrane domain insertase EMC complex, revealing an elongated cavity in the transmembrane region of the structure that can accommodate a weakly hydrophobic transmembrane helix. We have solved the structures of the protein N-glycosyltransferase (OST) and the protein O-mannosyltransferase Pmt1-Pmt2, revealing the evolutionarily conserved GT-C folds of their catalytic subunits. Protein trafficking requires lipid vesicle formation, a process that is dependent on lipid flippase activity to establish compositional asymmetry between the two leaflets of the bilayer. In this regard, we have determined the structures of all three classes of yeast lipid flippases. This renewal proposal continues our overarching goal to understand protein glycosylation and trafficking. We propose to address two specific knowledge gaps: the structure and mechanism of two protein mannosyltransferases, and how the recently discovered ternary protein complex Arl1-Gea2-Drs2 couples lipid flipping activity with membrane curvature formation, thereby facilitating the downstream vesicle budding process for protein and membrane trafficking. Drs2 is a phosphatidylserine flippase required for the formation of AP-1/clathrin-coated vesicles that travel back and forth between the trans-Golgi network (TGN) and early endosomes. The small GTPase Arl1 is a member of the ADP-ribosylation factor (Arf) family that is activated by the Arf guanine nucleotide exchange factor Gea2. Arl1 operates exclusively in the TGN and is the least well-understood member of the Arf family. We will reconstitute the ternary complex in vitro and perform a comprehensive structure-function study. Protein glycosylation and trafficking is intimately linked to tumorigenesis and cancer progression. Our mechanistic studies will fill these fundamental knowledge gaps, and our derived structures may facilitate the development of small molecules for cancer treatment.

项目摘要该建议的目的是获得对蛋白质糖基化的机械和病理理解和贩运。在上一个融资周期中，我们为这一过程做出了重要贡献。我们已经确定了跨膜结构域插入酶EMC复合物的结构，揭示了一个细长结构的跨膜区域的空腔，可以容纳弱疏水跨膜螺旋。我们已经解决了蛋白质N-糖基转移酶（OST）和蛋白O-甘露糖基转移酶PMT1-PMT2，揭示了其催化的进化保守的GT-C折叠亚基。蛋白质运输需要脂质囊泡形成，这一过程取决于脂质氟脂酶活性在双层的两个传单之间建立组成不对称。在这方面，我们有确定了所有三类酵母脂质氟）的结构。这项续签提案继续我们了解蛋白质糖基化和贩运的总体目标。我们建议解决两个特定的知识差距：两种蛋白质甘露糖基转移酶的结构和机制，以及最近如何发现三元蛋白复合物ARL1-GEA2-DRS2夫妇脂质翻转活性与膜曲率形成，从而促进了蛋白质和膜运输的下游囊泡发芽过程。 DRS2是形成AP-1/网格蛋白涂层的囊泡所需的磷脂酰丝氨酸丝。反式高尔基网络（TGN）和早期内体之间来回来回。小的GTPase ARL1是 ARF鸟嘌呤核苷酸交换激活的ADP-核糖基化因子（ARF）家族的成员因子GEA2。 ARL1专门在TGN中运作，是ARF家族中最不受欢迎的成员。我们将在体外重建三元复合物，并进行全面的结构功能研究。蛋白质糖基化和运输与肿瘤发生和癌症的进展密切相关。我们的机械研究将填补这些基本的知识差距，我们的衍生结构可能有助于发展用于癌症治疗的小分子。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cryo-EM structures of the endoplasmic reticulum membrane complex.

DOI：
10.1111/febs.15786
发表时间：
2022-01
期刊：
The FEBS journal
影响因子：
0
作者：
Bai L;Li H
通讯作者：
Li H

Structure of the human GlcNAc-1-phosphotransferase αβ subunits reveals regulatory mechanism for lysosomal enzyme glycan phosphorylation.

DOI：
10.1038/s41594-022-00748-0
发表时间：
2022-04
期刊：
NATURE STRUCTURAL & MOLECULAR BIOLOGY
影响因子：
16.8
作者：
Li, Hua;Lee, Wang-Sik;Feng, Xiang;Bai, Lin;Jennings, Benjamin C.;Liu, Lin;Doray, Balraj;Canfield, William M.;Kornfeld, Stuart;Li, Huilin
通讯作者：
Li, Huilin

Protein N-glycosylation and O-mannosylation are catalyzed by two evolutionarily related GT-C glycosyltransferases.