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Three-Dimensional Molecular Tracking of IgE-Fc{epsilon}RI in Live Cells

Award Number: R01AI097154
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 08/01/2012
PERIOD OF PERFORMANCE END DATE: 07/31/2017

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Three-Dimensional Molecular Tracking of IgE-Fc{epsilon}RI in Live Cells - DESCRIPTION (provided by applicant): This work substantially expands the technical capabilities of newly developed three-dimensional molecular tracking methods. These technical advances will be exploited to study key protein-protein interactions important for the allergic response. This new approach to three dimensional molecular tracking uses four overlaping confocal volume elements and active feedback in X, Y, and Z once every 5 milliseconds to follow 3D dynamic molecular motion inside of live cells. As a model system, we propose to follow the 3D spatio- temporal dynamics of Fc?RI-IgE. As the key mediator for the allergic responses, this system has significant relevance to human health. Tracking of this high affinity IgE receptor is enabled through the use of IgE- quantum dot (QD) probes. Advantages of our approach to 3D molecular tracking include the ability to follow QDs in high background environments and the ability to perform time-resolved spectroscopy on the molecules while they are being followed. Here, we propose to significantly improve the contextual information available and the spatio-temporal resolution of this newly developed 3D tracking method. Moreover, these substantial technical improvements will be used to explore crucial steps involved in Fc?RI-IgE signaling and down- regulation via endocytosis. The specific aims of this proposal are: Aim 1. To increase the contextual and spatio-temporal resolution of the 3D tracking microscope. This Aim includes methods to simultaneous image key GFP labeled complexes during 3D molecular tracking of QD labeled Fc?RI-IgE, the acquisition of longer 3D trajectories using blinking-suppressed quantum dots, and methods to increase the spatio-temporal resolution of our 3D tracking methods. Aim 2. To apply these new advances to the analysis of molecular mechanisms that govern trafficking and signaling competency of internalized Fc?RI. This Aim explores the spatio-temporal dynamics of Fc?RI-IgE complexes interacting with actin, mircotubules, clathrin, and the kinases Lyn and Syk.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2018 ( Subtotal = -$92,327 )
2018	2015	LOS ALAMOS NATIONAL SECURITY, LLC	BIKINI ATOLL RD SM 30	LOS ALAMOS	NM	87545	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	4	1/17/2018	NON-COMPETING CONTINUATION	-$92,327
														Subtotal = -$92,327

Issue Date FY: 2015 ( Subtotal = $407,018 )
2015	2015	LOS ALAMOS NATIONAL SECURITY LLC	4200 WEST JEMEZ ROAD	LOS ALAMOS	NM	87544	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	4	6/30/2015	NON-COMPETING CONTINUATION	$407,018
														Subtotal = $407,018

Issue Date FY: 2014 ( Subtotal = $407,996 )
2014	2014	LOS ALAMOS NATIONAL SECURITY LLC	4200 WEST JEMEZ ROAD	LOS ALAMOS	NM	87544	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	3	7/8/2014	NON-COMPETING CONTINUATION	$407,996
														Subtotal = $407,996

Issue Date FY: 2013 ( Subtotal = $382,484 )
2013	2013	LOS ALAMOS NATIONAL SECURITY LLC	4200 WEST JEMEZ ROAD	LOS ALAMOS	NM	87544	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	2	6/26/2013	NON-COMPETING CONTINUATION	$382,484
														Subtotal = $382,484

Issue Date FY: 2012 ( Subtotal = $405,833 )
2012	2012	LOS ALAMOS NATIONAL SECURITY LLC	4200 WEST JEMEZ ROAD	LOS ALAMOS	NM	87544	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	1	8/1/2012	NEW	$405,833
														Subtotal = $405,833

Grand Total All Awards = $1,511,004

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Three-Dimensional Molecular Tracking of IgE-Fc{epsilon}RI in Live Cells

Award Number: R01AI097154

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 08/01/2012

PERIOD OF PERFORMANCE END DATE: 07/31/2017

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