Proteins that behave as switches hold great promise as selective protein therapeutics. Previous studies by the PI have demonstrated that protein switches can be engineered by creating a large number of different fusions between two genes and subjecting these gene fusions to evolution in the lab. Such directed evolution strategies will be used to develop and improve protein switches to effectively activate prodrugs selectively in cancer cells. These switches will be evaluated for their ability to eliminate tumor xenographs in vivo using endogenous expression and via systemic delivery. These switches in combination with the appropriate prodrug have potential as a targeted therapeutic for the treatment of cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM066972-10
Application #
8503846
Study Section
Macromolecular Structure and Function B Study Section (MSFB)
Program Officer
Gerratana, Barbara
Project Start
2003-07-01
Project End
2017-02-28
Budget Start
2013-04-01
Budget End
2014-02-28
Support Year
10
Fiscal Year
2013
Total Cost
$309,134
Indirect Cost
$101,634
Name
Johns Hopkins University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Valdes, Gilmer; Schulte, Reinhard W; Ostermeier, Marc et al. (2014) The High-Affinity Maltose Switch MBP317-347 has Low Affinity for Glucose: Implications for Targeting Tumors with Metabolically Directed Enzyme Prodrug Therapy. Chem Biol Drug Des 83:266-71
Chaikind, Brian; Ostermeier, Marc (2014) Directed evolution of improved zinc finger methyltransferases. PLoS One 9:e96931
Kanwar, Manu; Wright, R Clay; Date, Amol et al. (2013) Protein switch engineering by domain insertion. Methods Enzymol 523:369-88
Guntas, Gurkan; Kanwar, Manu; Ostermeier, Marc (2012) Circular permutation in the ýý-loop of TEM-1 ýý-lactamase results in improved activity and altered substrate specificity. PLoS One 7:e35998
Cheung, Luthur Siu-Lun; Kanwar, Manu; Ostermeier, Marc et al. (2012) A hot-spot motif characterizes the interface between a designed ankyrin-repeat protein and its target ligand. Biophys J 102:407-16
Tullman, Jennifer; Guntas, Gurkan; Dumont, Matthew et al. (2011) Protein switches identified from diverse insertion libraries created using S1 nuclease digestion of supercoiled-form plasmid DNA. Biotechnol Bioeng 108:2535-43
Wright, Chapman M; Wright, R Clay; Eshleman, James R et al. (2011) A protein therapeutic modality founded on molecular regulation. Proc Natl Acad Sci U S A 108:16206-11
Heins, Richard A; Choi, Jay H; Sohka, Takayuki et al. (2011) In vitro recombination of non-homologous genes can result in gene fusions that confer a switching phenotype to cells. PLoS One 6:e27302
Ostermeier, Marc (2009) Designing switchable enzymes. Curr Opin Struct Biol 19:442-8
Berrondo, Monica; Ostermeier, Marc; Gray, Jeffrey J (2008) Structure prediction of domain insertion proteins from structures of individual domains. Structure 16:513-27