PROTEINS REGULATING CU-ATPASES IN EPITHELIA
PROTEINS REGULATING CU-ATPASES IN EPITHELIA
批准号:
7690603
负责人:
Ann L Hubbard
金额:
$29.58万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2009
资助国家:
美国
项目状态:
已结题
起止时间:
2009-09-15 至 2014-08-31
关键词:
ATP phosphohydrolaseAffectAmino AcidsAnimalsApicalB-LymphocytesBile fluidBindingBinding SitesBiochemicalBiologyBiotinylationBlood CirculationBreedingC-terminalCaco-2 CellsCell membraneCell surfaceCellsChimera organismCopperCytoplasmic ProteinCytoplasmic VesiclesEnvironmentEpithelial CellsEpitheliumExcisionGoalsHepaticHepatocyteHeterozygoteHomeostasisHomoHumanHybridsImmunofluorescence ImmunologicIn VitroIntestinesIonsKidneyKnock-outLengthLettersLiverMDCK cellMediatingMembraneMetalsMethodsMicrotubulesMolecularMolecular ChaperonesMolecular ConformationMovementMusN-terminalPDZ proteinPathway interactionsPlayProtein BindingProteinsRegulationRoleRouteSignal TransductionSorting - Cell MovementTacrolimus Binding ProteinsTailTestingTissuesTitrationsTyrosineVesicleWilson disease proteinWorkYeastsapical membranebasebathocuproine disulfonatedimerdynactinhuman diseasein vivokidney cellknock-downmutantnoveloverexpressionpolarized cellprotein protein interactiontacrolimus binding protein 4traffickingtrans-Golgi Networkyeast two hybrid system
中文摘要
我们的长期目标是阐明两种哺乳动物cu - atp酶的分子机制
英文摘要
Our long-term goal is to elucidate the molecular mechanisms by which two mammalian Cu-ATPases
regulate copper homeostasis in polarized epithelial cells. These Cu-ATPases have two functions that
necessitate their intracellular redistribution, or trafficking, in a copper-sensitive and reversible manner. First,
they transport copper into the secretory pathway to metallate newly-synthesized cuproenzymes. They also
efflux copper. ATP7A in intestinal epithelial cells delivers dietary Cu to the circulation (basolateral
environment), and ATP7B in hepatic cells delivers excess Cu to the bile (apical environment). This project
focuses on cellular proteins that modulate the Cu-ATPases' trafficking and export functions. In Aim 1 A, we
will determine the dynamics of ATP7B in the polarized hepatic WIF-B cells, after over-expressing or knocking
down the dynactin subunit, p62, which binds the N-terminal domain of ATP7B (not ATP7A) when copper is
elevated. p62 is proposed to direct the apical movement of ATP7B-positive vesicles along microtubules.
Titration of cellular copper with immunofluorescence and copper-64 efflux determinations will be performed.
Aim 1B focuses on a 9 amino acid apical targeting motif we discovered in the N-terminus of only ATP7B. We
will collaborate with Projects 3 + 4 to determine if the targeting motif interacts with other domains of ATP7B
and if a chimera containing this motif fused to a basolateral protein will target to the apical membrane. To
identify the protein (X) proposed to bind the targeting motif, we will use 2 approaches: a membrane-based
yeast-two hybrid system; and a screen of putative binding domains of candidate proteins. In Aim 2, we will
determine the mechanism by which a PDZ protein, AIPP, targets/retains endogenous ATP7A to/in the
basolateral region of polarized intestinal Caco-2 cells. We will over-express and knock down AIPP1 using
approaches similar to those in Aim 1A and collaborate with Project 3 to determine if/how AIPP1 levels affect
Cu export. In Aim 3, we will study the role of ATOX1, the copper chaperone for both Cu-ATPases, and its
proposed modifier, FKBP52, in copper homeostasis in vivo in compound heterozygote mice (ATOX1+/-;
FKBP+/-). We will collaborate with Projects 2 and 3 to study the liver, intestine and kidney in these animals.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Imaging Core
-
批准号:8012349
-
项目类别:
-
资助金额:$18.86万
-
财政年份:2011
-
负责人:Ann L Hubbard
-
依托单位:
Cu Homeostasis and Cu-ATPases in Polarized Epithelia
-
批准号:7487972
-
项目类别:
-
资助金额:$29.25万
-
财政年份:2007
-
负责人:Ann L Hubbard
-
依托单位:
Cu Homeostasis and Cu-ATPases in Polarized Epithelia
-
批准号:7133531
-
项目类别:
-
资助金额:$31.37万
-
财政年份:2006
-
负责人:Ann L Hubbard
-
依托单位:
AMT DUAL CAMERA SYST: PARASITES: PLASMODIUM, C FASCICULATA, T BRUCEI
-
批准号:7166513
-
项目类别:
-
资助金额:$2.84万
-
财政年份:2005
-
负责人:Ann L Hubbard
-
依托单位:
AMT DUAL CAMERA: WILSON DIS PROTEIN, CORONAVIRUS & INFECTIOUS BRONCHITIS VIRUS
-
批准号:7166511
-
项目类别:
-
资助金额:$2.84万
-
财政年份:2005
-
负责人:Ann L Hubbard
-
依托单位:
AMT DUAL CAMERA: ID GENE & PROTEINS IN BRAIN DVMT & RETINAL PIGMENTED EPITHELIUM
-
批准号:7166512
-
项目类别:
-
资助金额:$2.84万
-
财政年份:2005
-
负责人:Ann L Hubbard
-
依托单位:
AMT Dual Camera System
-
批准号:6876414
-
项目类别:
-
资助金额:$14.22万
-
财政年份:2005
-
负责人:Ann L Hubbard
-
依托单位:
AMT DUAL CAMERA SYST: BACTERIOLOGY: MYXOCOCCUS XANTHUS, YERSINA ENTEROCOLITICA
-
批准号:7166510
-
项目类别:
-
资助金额:$2.84万
-
财政年份:2005
-
负责人:Ann L Hubbard
-
依托单位:
AMT DUAL CAMERA SYST: GENOMICS & PROTEOMICS, WOUND RE-EPITHELIALIZATION
-
批准号:7166509
-
项目类别:
-
资助金额:$2.84万
-
财政年份:2005
-
负责人:Ann L Hubbard
-
依托单位:
Development of Polarity in Hepatic Cells
-
批准号:7110343
-
项目类别:
-
资助金额:$41.79万
-
财政年份:2004
-
负责人:Ann L Hubbard
-
依托单位:
Development of Polarity in Hepatic Cells
-
批准号:6952811
-
项目类别:
-
资助金额:$41.41万
-
财政年份:2004
-
负责人:Ann L Hubbard
-
依托单位:
Development of Polarity in Hepatic Cells
-
批准号:7483700
-
项目类别:
-
资助金额:$42.38万
-
财政年份:2004
-
负责人:Ann L Hubbard
-
依托单位:
Development of Polarity in Hepatic Cells
-
批准号:7277594
-
项目类别:
-
资助金额:$41.89万
-
财政年份:2004
-
负责人:Ann L Hubbard
-
依托单位:
Development of Polarity in Hepatic Cells
-
批准号:6824213
-
项目类别:
-
资助金额:$51.27万
-
财政年份:2004
-
负责人:Ann L Hubbard
-
依托单位:
MEMBRANE DYNAMICS IN POLARIZED EPITHELIAL CELLS
-
批准号:6564273
-
项目类别:
-
资助金额:$14.67万
-
财政年份:2002
-
负责人:Ann L Hubbard
-
依托单位:
Plasma Membrane Trafficking in Polarized Hepatocytes
-
批准号:6420652
-
项目类别:
-
资助金额:$34.66万
-
财政年份:2002
-
负责人:Ann L Hubbard
-
依托单位:
Plasma Membrane Trafficking in Polarized Hepatocytes
-
批准号:6724849
-
项目类别:
-
资助金额:$34.66万
-
财政年份:2002
-
负责人:Ann L Hubbard
-
依托单位:
Plasma Membrane Trafficking in Polarized Hepatocytes
-
批准号:6878112
-
项目类别:
-
资助金额:$34.66万
-
财政年份:2002
-
负责人:Ann L Hubbard
-
依托单位:
CORE--CELL CULTURE /TRANSFECTION
-
批准号:6564276
-
项目类别:
-
资助金额:$14.67万
-
财政年份:2002
-
负责人:Ann L Hubbard
-
依托单位:
Plasma Membrane Trafficking in Polarized Hepatocytes
-
批准号:6620691
-
项目类别:
-
资助金额:$34.66万
-
财政年份:2002
-
负责人:Ann L Hubbard
-
依托单位:
海外基金