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 #
5R01GM095930-02
Application #
8197577
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
2011-12-01
Budget End
2012-11-30
Support Year
2
Fiscal Year
2012
Total Cost
$323,580
Indirect Cost
$123,580
Name
Johns Hopkins University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
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Sarabipour, Sarvenaz; King, Christopher; Hristova, Kalina (2014) Uninduced high-yield bacterial expression of fluorescent proteins. Anal Biochem 449:155-7
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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
Sarabipour, Sarvenaz; Hristova, Kalina (2013) Glycophorin A transmembrane domain dimerization in plasma membrane vesicles derived from CHO, HEK 293T, and A431 cells. Biochim Biophys Acta 1828:1829-33
Li, Edwin; Wimley, William C; Hristova, Kalina (2012) Transmembrane helix dimerization: beyond the search for sequence motifs. Biochim Biophys Acta 1818:183-93
He, Lijuan; Shobnam, Nadia; Wimley, William C et al. (2011) FGFR3 heterodimerization in achondroplasia, the most common form of human dwarfism. J Biol Chem 286:13272-81
Marks, Jessica R; Placone, Jesse; Hristova, Kalina et al. (2011) Spontaneous membrane-translocating peptides by orthogonal high-throughput screening. J Am Chem Soc 133:8995-9004
Wimley, William C; Hristova, Kalina (2011) Antimicrobial peptides: successes, challenges and unanswered questions. J Membr Biol 239:27-34