QI-FRET: a new tool in Receptor Tyrosine Kinase research Kalina Hristova, Johns Hopkins University Ligand-independent homo and heterodimerization of receptor tyrosine kinases (RTKs) is a process that is not well understood despite being implicated in various human pathologies. The lack of knowledge is mainly due to a lack of an appropriate methodology to measure the dimerization energetics of full-length RTK. The lack of knowledge is, in turn, a bottleneck in developing effective RTK-targeted therapies. Here we propose to (i) develop a quantitative imaging FRET (QI-FRET) method that yields dimerization free energies in plasma membrane-derived vesicles and to (ii) characterize the homo and heterodimerization free energies of the ErbB and FGF receptors, chosen because of their very strong link to human disease. Upon the completion of this work, we will be able to predict the degree of ligand-independent dimerization, and thus biological activity, as a function of RTK expression. The method that will be established and the knowledge gained will ultimately aid the search and refinement of effective RTK-targeted treatments for cancers and growth disorders.

Public Health Relevance

We propose to (i) develop a quantitative imaging FRET (QI-FRET) method that yields dimerization free energies in plasma membrane-derived vesicles and to (ii) characterize the homo and heterodimerization free energies of the ErbB and FGF receptors, chosen because of their very strong link to human disease. Upon the completion of this work, we will be able to predict the degree of ligand-independent dimerization, and thus biological activity, as a function of RTK expression. The method that will be established and the knowledge gained will ultimately aid the search and refinement of effective RTK-targeted treatments for cancers and growth disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM095930-04
Application #
8588339
Study Section
Special Emphasis Panel (ZRG1-BCMB-M (90))
Program Officer
Chin, Jean
Project Start
2010-12-01
Project End
2014-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
4
Fiscal Year
2014
Total Cost
$290,814
Indirect Cost
$110,814
Name
Johns Hopkins University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Sarabipour, Sarvenaz; Hristova, Kalina (2016) Mechanism of FGF receptor dimerization and activation. Nat Commun 7:10262
Sarabipour, Sarvenaz; Ballmer-Hofer, Kurt; Hristova, Kalina (2016) VEGFR-2 conformational switch in response to ligand binding. Elife 5:e13876
Wiedman, Gregory; Wimley, William C; Hristova, Kalina (2015) Testing the limits of rational design by engineering pH sensitivity into membrane-active peptides. Biochim Biophys Acta 1848:951-7
Sarabipour, Sarvenaz; Chan, Robin B; Zhou, Bowen et al. (2015) Analytical characterization of plasma membrane-derived vesicles produced via osmotic and chemical vesiculation. Biochim Biophys Acta 1848:1591-8
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
Placone, Jesse; He, Lijuan; Del Piccolo, Nuala et al. (2014) Strong dimerization of wild-type ErbB2/Neu transmembrane domain and the oncogenic Val664Glu mutant in mammalian plasma membranes. Biochim Biophys Acta 1838:2326-30
Kavran, Jennifer M; McCabe, Jacqueline M; Byrne, Patrick O et al. (2014) How IGF-1 activates its receptor. Elife 3:
Sarabipour, Sarvenaz; King, Christopher; Hristova, Kalina (2014) Uninduced high-yield bacterial expression of fluorescent proteins. Anal Biochem 449:155-7

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