We will determine the contributions of the four receptor tyrosine kinase (RTK) domains to the energetics of RTK lateral dimerization. The six receptors chosen for this study, ErbB1, ErbB2, ErbB3, FGFR1, FGFR2, and FGFR3, have been linked to human pathologies. The contributions of the different domains in these six RTKs to the dimerization free energies are currently unknown, mainly due to experimental challenges in the study of full-length RTKs, and membrane proteins in general. Here we will use a novel experimental approach, based on quantitative FRET, which yields dimerization thermodynamics in plasma membranes derived from mammalian cells. The work proposed here is the first step towards comprehensive characterization of the interplay between the different RTK domains in signaling. It will provide basic knowledge regarding the role of the different domains in the dimerization process, and thus aid in the development of highly specific therapeutics which can be used to treat cancers and growth disorders.

Public Health Relevance

RTK domains and RTK dimerization thermodynamics Narrative Many pathologies are believed to occur due to disregulation of ligand-independent RTK dimerization. Here we will gain insight into this process by determining the contributions of the four receptor tyrosine kinase (RTK) domains to the energetics of RTK dimerization. The proposed work will aid in the development of highly specific therapeutics that can be used to treat cancers and growth disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM068619-09
Application #
8536822
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Chin, Jean
Project Start
2004-05-01
Project End
2014-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
9
Fiscal Year
2013
Total Cost
$308,700
Indirect Cost
$117,630
Name
Johns Hopkins University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
King, Christopher; Stoneman, Michael; Raicu, Valerica et al. (2016) Fully quantified spectral imaging reveals in vivo membrane protein interactions. Integr Biol (Camb) 8:216-29
Sarabipour, Sarvenaz; Hristova, Kalina (2016) Effect of the achondroplasia mutation on FGFR3 dimerization and FGFR3 structural response to fgf1 and fgf2: A quantitative FRET study in osmotically derived plasma membrane vesicles. Biochim Biophys Acta 1858:1436-42
Sarabipour, Sarvenaz; Hristova, Kalina (2016) Mechanism of FGF receptor dimerization and activation. Nat Commun 7:10262
Singh, Deo R; Pasquale, Elena B; Hristova, Kalina (2016) A small peptide promotes EphA2 kinase-dependent signaling by stabilizing EphA2 dimers. Biochim Biophys Acta 1860:1922-8
Sarabipour, Sarvenaz; Ballmer-Hofer, Kurt; Hristova, Kalina (2016) VEGFR-2 conformational switch in response to ligand binding. Elife 5:e13876
Sarabipour, Sarvenaz; Hristova, Kalina (2015) FGFR3 unliganded dimer stabilization by the juxtamembrane domain. J Mol Biol 427:1705-14
Singh, Deo R; Ahmed, Fozia; King, Christopher et al. (2015) EphA2 Receptor Unliganded Dimers Suppress EphA2 Pro-tumorigenic Signaling. J Biol Chem 290:27271-9
Singh, Deo R; Cao, QingQing; King, Christopher et al. (2015) Unliganded EphA3 dimerization promoted by the SAM domain. Biochem J 471:101-9
Sarabipour, Sarvenaz; Del Piccolo, Nuala; Hristova, Kalina (2015) Characterization of membrane protein interactions in plasma membrane derived vesicles with quantitative imaging Förster resonance energy transfer. Acc Chem Res 48:2262-9
Del Piccolo, Nuala; Placone, Jesse; Hristova, Kalina (2015) Effect of thanatophoric dysplasia type I mutations on FGFR3 dimerization. Biophys J 108:272-8

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