Membrane properties of the OHC system
Membrane properties of the OHC system
批准号:
10408895
负责人:
JOSEPH R SANTOS-SACCHI
金额:
$16.71万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2017
资助国家:
美国
项目状态:
已结题
起止时间:
2017-06-12 至 2022-05-31
关键词:
AffectAnionsBinding SitesBiological AssayChargeContractsCysteineDataElevatorFamilyFamily memberFluorescence Resonance Energy TransferHearingHumanLabelMeasurableMeasuresMembraneMethodsModelingMolecularMolecular ConformationMotionMotorMovementOpticsPhysiologicalPropertyProteinsResolutionStructureSystemTransmembrane DomainWorkbasedata modelingdimerexperimental studyfluorophoremembermonomerprotein functionrat Pres proteinsingle moleculesingle-molecule FRETtool
中文摘要
项目总结
英文摘要
Project Summary
In this proposal we seek to develop single molecule FRET in prestin. The method will allow us to assess the
function of the protein using optical measures. Single molecule FRET has been developed in other proteins to
aid in the understanding of their function. Presently, we do not have a functional assay of prestin with a single
molecule resolution.
The rationale for the experiments is based on structural data interpolated from the structure of several
members of the extended anion transporter family to which prestin belongs. Available structural data together
with modeling data suggest that members of this family have 14 transmembrane domains with an inverted
repeat 7+7 structure to each monomer. The functional units of these proteins appear to be dimers. Each
monomer has a gate and core domain. We model prestin to behave like these transporters using an elevator
mechanism to move from contracted to expanded states. An analogous movement in these family members
from inside open to outside open conformations result in a relative motion between two parts of the same
monomer. Thus, there is a movement of the gate domain (Tm domains 5-8 and 12-14) relative to the stable
core domain (Tm domains 1-4 and 9-11) containing the Tm anion binding site. Based on these models we will
insert several cysteine residues in the gate domain that will allow us to label a single dimer with two different
fluorophores with FRET activity. We have already generated a cysteine free protein that shows reduced but
measurable gating charge movement (NLC), confirming its functionality. The distance between the
fluorophores is modelled to vary by 30A0 that would give rise to a significant change in FRET efficiency.
Establishing a single molecule FRET assay will allow us to measure how physiological parameters affect the
function of the protein.
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会议论文
Membrane properties of the OHC system
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批准号:9364111
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项目类别:
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资助金额:$64.77万
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财政年份:2017
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Membrane properties of the OHC system
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批准号:10163155
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项目类别:
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资助金额:$64.97万
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财政年份:2017
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Structural correlates of prestin activity.
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批准号:7333293
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项目类别:
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资助金额:$34.66万
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财政年份:2007
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Structural correlates of prestin activity.
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批准号:7546556
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项目类别:
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资助金额:$34.71万
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财政年份:2007
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Structural correlates of prestin activity.
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批准号:7213152
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项目类别:
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资助金额:$35.06万
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财政年份:2007
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Membrane properties of cells comprising the OHC system
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批准号:7157225
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项目类别:
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资助金额:$4.51万
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财政年份:2005
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
IN VIVO ANALYSIS OF OUTER HAIR CELL MOTILITY
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批准号:2127082
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项目类别:
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资助金额:$13.12万
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财政年份:1993
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
IN VIVO ANALYSIS OF OUTER HAIR CELL MOTILITY
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批准号:2127081
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项目类别:
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资助金额:$13.05万
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财政年份:1993
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
IN VIVO ANALYSIS OF OUTER HAIR CELL MOTILITY
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批准号:3218536
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项目类别:
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资助金额:$13.28万
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财政年份:1993
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
CELL COUPLING IN THE INNER EAR
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批准号:3072031
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项目类别:
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资助金额:$5.33万
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财政年份:1985
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
CELL COUPLING IN THE INNER EAR
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批准号:3074833
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项目类别:
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资助金额:$5.27万
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财政年份:1985
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
CELL COUPLING IN THE INNER EAR
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批准号:3074831
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项目类别:
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资助金额:$5.4万
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财政年份:1985
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
CELL COUPLING IN THE INNER EAR
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批准号:3074832
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项目类别:
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资助金额:$5.32万
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财政年份:1985
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
CELL COUPLING IN THE INNER EAR
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批准号:3074830
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项目类别:
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资助金额:$5.44万
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财政年份:1985
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Membrane properties of cells comprising the OHC system
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批准号:7074896
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项目类别:
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资助金额:$2.2万
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财政年份:1984
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
MEMBRANES PROPERTIES OF CELLS COMPRISING THE OHC SYSTEM
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批准号:3216385
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项目类别:
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资助金额:$15.52万
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财政年份:1984
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Membrane properties of cells comprising the OHC system
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批准号:7385129
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项目类别:
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资助金额:$46.06万
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财政年份:1984
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Membrane properties of cells comprising the OHC system
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批准号:8248686
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项目类别:
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资助金额:$42.18万
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财政年份:1984
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
Membrane properties of cells comprising the OHC system
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批准号:8643196
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项目类别:
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资助金额:$42.18万
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财政年份:1984
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
CELL-TO-CELL COMMUNICATION IN THE ORGAN OF CORTI
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批准号:3402483
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项目类别:
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资助金额:$3.5万
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财政年份:1984
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负责人:JOSEPH R SANTOS-SACCHI
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依托单位:
海外基金