Molecular basis of congenital disorder of glycosylation type 1N
Molecular basis of congenital disorder of glycosylation type 1N
批准号:
10700974
负责人:
ANANT K MENON
金额:
$21.19万
依托单位国家:
美国
项目类别:
财政年份:
2022
资助国家:
美国
项目状态:
已结题
起止时间:
2022-09-08 至 2024-08-31
关键词:
BindingBinding ProteinsBlindnessCadherinsCell Adhesion MoleculesCell Surface ReceptorsCell-Free SystemCellsChemicalsChildClinicalCollaborationsCongenital disorders of glycosylationCryoelectron MicroscopyCytoplasmDefectDevelopmentDevelopmental Delay DisordersDiseaseDolicholEndoplasmic ReticulumEnzymesEpitopesEuropeExtracellular Matrix ProteinsFaceFailure to ThriveFamilyFibroblastsGenetic DiseasesGlycoproteinsGoalsHuman GeneticsIndiaIntegrinsLamininLateralLifeLinkLipidsLocationMapsMembraneMembrane BiologyMembrane ProteinsMetabolic DiseasesMicrocephalyMicrosomesMiddle EastMissense MutationModelingMolecularMorphologyMuscle hypotoniaNational Institute of Child Health and Human DevelopmentNatureNorth AmericaOligosaccharidesOrganogenesisPathway interactionsPatientsPermeabilityPhenotypePolysaccharidesPropertyProteinsRoleSeizuresSensorineural Hearing LossSideSiteSpeech DelayStructureSymptomsTestingTherapeuticTissuesUnited States National Institutes of HealthVesicleX-Ray Crystallographyautosomecognitive disabilityexperienceexperimental studyglycosylationglycosyltransferaseimprovedinterestisoprenoidnovel strategiesphysically handicappedpreventprogramssealstroke-like episodesugartherapy developmentthree dimensional structuretreatment strategy
中文摘要
N-糖基化是生命所必需的。钙粘附素、整合素和层粘连蛋白等糖蛋白是
发育、器官发生和组织结构。N-糖基化缺陷是许多人类
遗传性疾病包括一组异质性常染色体隐性遗传代谢性疾病,称为
先天性糖基化紊乱(CDGs)。CDG出现了一系列毁灭性的症状,包括
发育迟缓、发育迟缓、视力丧失、低眼压、小头畸形、癫痫发作和中风--
就像小插曲。患有CDG的儿童患有认知和身体残疾。虽然这些症状中的一些可能
如果接受治疗,慢性萎缩性胃炎无法治愈。
我们在R21应用中的目标是探索CDG 1N的分子基础,CDG 1N是一种知之甚少的
由内质网膜蛋白RFT1错义突变引起的疾病。病人们,
到目前为止在北美、英国/欧洲、中东和印度都发现了一种独特的
除了典型的严重的CDG症状外,还有感觉神经性耳聋。CDG1N目前还没有治疗方法。不像
大多数CDG的潜在分子缺陷被很好地理解,例如
糖基转移酶或糖加工酶CDG1N是一个谜,因为RFT1的功能不是
为人所知。在CDG1N细胞中,N-糖基化途径在关键阶段被阻断,导致
一种不能被糖基化机制利用的脂质中间体(称为M5-DLO)。我们假设
RFT1在细胞中的关键作用是催化利用M5-DLO,无论是通过克服障碍
由内质网结构/形态构成,以便于其移交给下游机械,或
通过防止其错误本地化。
我们将在两个具体目标上检验这一假设。第一个目标将集中在RFT1在M5-DLO中的作用
利用RFT1缺陷细胞(包括CDG1N患者成纤维细胞)、无细胞系统和
内质网细分的新方法。我们还将测试RFT1是否为M5-DLO
结合蛋白。认识到结构信息对于理解RFT1的功能至关重要
在分子水平上,第二个目标是建立RFT1的结构-功能模型。在这里,我们将定义其
膜拓扑结构,确定关键功能残基,并启动一个程序来阐明其三维结构
结构。
从长远来看,我们的结果将解释为什么RFT1缺陷会导致M5-DLO积聚和
因此,CDG1N,从而为制定治疗战略和治疗方法铺平了道路
可替代RFT1‘S功能,恢复糖基化,改善临床症状。
英文摘要
N-glycosylation is essential for life. Glycoproteins such as cadherins, integrins and laminins are key to
development, organogenesis, and tissue organization. Defects in N-glycosylation underlie numerous human
genetic disorders including a heterogeneous group of autosomal-recessive, metabolic diseases termed
Congenital Disorders of Glycosylation (CDGs). CDGs present with a range of devastating symptoms, including
failure to thrive, developmental and speech delays, vision loss, hypotonia, microcephaly, seizures, and stroke-
like episodes. Children with CDGs suffer cognitive and physical disabilities. While some of these symptoms can
be treated, there is no cure for CDGs.
Our goal in this R21 application is to explore the molecular basis of CDG type 1N, a poorly understood
disease caused by missense mutations in the endoplasmic reticulum membrane protein RFT1. Patients,
identified thus far in North America, U.K./Europe, the Middle East, and India, often present with a unique
sensorineural deafness in addition to typical severe CDG symptoms. There is no therapy for CDG1N. Unlike
most CDGs for which the underlying molecular defect is well understood, e.g., deficiency of a
glycosyltransferase or sugar-processing enzyme, CDG1N is an enigma because the function of RFT1 is not
known. The pathway of N-glycosylation is blocked at a critical stage in CDG1N cells, resulting in the build-up of
a lipid intermediate (termed M5-DLO) that cannot be utilized by the glycosylation machinery. We hypothesize
that the critical role of RFT1 in cells is to catalyze utilization of M5-DLO either by overcoming an impediment
posed by endoplasmic reticulum structure/morphology to facilitate its handoff to downstream machinery, or
by preventing its mis-localization.
We will test this hypothesis in two specific aims. The first aim will focus on the role of RFT1 in M5-DLO
utilization, making use of RFT1-deficient cells (including CDG1N patient fibroblasts), cell-free systems, and a
novel approach to sub-fractionating the endoplasmic reticulum. We will also test whether RFT1 is an M5-DLO
binding protein. Recognizing that structural information is critical to understand the function of RFT1 at a
molecular level, the second aim will develop a structure-function model of RFT1. Here we will define its
membrane topology, identify key functional residues, and initiate a program to elucidate its 3-dimensional
structure.
In the long term, our results will illuminate why RFT1 deficiency results in M5-DLO accumulation and
consequently CDG1N, thereby paving the way for the development of treatment strategies and therapeutics that
could subserve RFT1's function to restore glycosylation and improve clinical symptoms.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Scramblases for protein glycosylation
-
批准号:10420706
-
项目类别:
-
资助金额:$56.34万
-
财政年份:2022
-
负责人:ANANT K MENON
-
依托单位:
Molecular basis of congenital disorder of glycosylation type 1N
-
批准号:10510784
-
项目类别:
-
资助金额:$25.43万
-
财政年份:2022
-
负责人:ANANT K MENON
-
依托单位:
Scramblases for protein glycosylation
-
批准号:10600063
-
项目类别:
-
资助金额:$51.2万
-
财政年份:2022
-
负责人:ANANT K MENON
-
依托单位:
Rhodopsin-mediated phospholipid flipping
-
批准号:8786659
-
项目类别:
-
资助金额:$25.43万
-
财政年份:2014
-
负责人:ANANT K MENON
-
依托单位:
Rhodopsin-mediated phospholipid flipping
-
批准号:8895952
-
项目类别:
-
资助金额:$20.76万
-
财政年份:2014
-
负责人:ANANT K MENON
-
依托单位:
Structural Analysis of the GPI Transamidase Complex
-
批准号:8267601
-
项目类别:
-
资助金额:$21.13万
-
财政年份:2011
-
负责人:ANANT K MENON
-
依托单位:
Structural Analysis of the GPI Transamidase Complex
-
批准号:8196655
-
项目类别:
-
资助金额:$25.35万
-
财政年份:2011
-
负责人:ANANT K MENON
-
依托单位:
Biosynthesis of Membrane Protein Glycolipid Anchors
-
批准号:7938503
-
项目类别:
-
资助金额:$19.8万
-
财政年份:2009
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:7080488
-
项目类别:
-
资助金额:$31.17万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:7255834
-
项目类别:
-
资助金额:$30.27万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:7495987
-
项目类别:
-
资助金额:$30.27万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:8389633
-
项目类别:
-
资助金额:$48.85万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:7783154
-
项目类别:
-
资助金额:$35.77万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:6929555
-
项目类别:
-
资助金额:$31.92万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:8010138
-
项目类别:
-
资助金额:$35.41万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:8197567
-
项目类别:
-
资助金额:$35.41万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid Flip-flop in Biogenic Membranes
-
批准号:8529808
-
项目类别:
-
资助金额:$4.98万
-
财政年份:2005
-
负责人:ANANT K MENON
-
依托单位:
FASEB Conference-Protein Lipidation and Membrane Domains
-
批准号:6808660
-
项目类别:
-
资助金额:$0.65万
-
财政年份:2004
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid flip-flop in biogenic membranes
-
批准号:6520496
-
项目类别:
-
资助金额:$10.91万
-
财政年份:2001
-
负责人:ANANT K MENON
-
依托单位:
Phospholipid flip-flop in biogenic membranes
-
批准号:6319553
-
项目类别:
-
资助金额:$10.91万
-
财政年份:2001
-
负责人:ANANT K MENON
-
依托单位:
海外基金