Covalent modification by isoprenoid lipids (prenylation) is a critical post-translational event for many proteins involved in cellular signaling and cancer. The primary goal of this research program is to design and test prenyl function inhibitors and to identify and characterize proteins that are farnesylated in vivo. The studies outlined in this proposal will result in the preparation of new tools to probe the specificity of the prenyltransferases protein- farnesyltransferase (FTase) and protein-geranylgeranyltransferase (GGTase-I), enable further development of prenyl function inhibitors and identify new prenylated proteins as potential targets for therapeutic intervention. By varying the chemical structure of the prenyl lipid, we are developing reagents to probe the biological function of the posttranslational modification. Taking advantage of the fact that the prenyl group forms a substantial part of the peptide substrate binding site in the prenyltransferases has allowed us to develop peptide selective inhibitors of prenyl function. These unnatural analogues may allow for the selective interference with specific prenylation targets and may provide lead compounds to alleviate the potential toxicity associated with complete inhibition of protein prenylation. In particular, the unnatural analogues may be useful to obtain a more complete understanding of the role that alternative prenylation plays in Farnesyl transferase inhibitor (FTI) evasion by oncogenes such as K-Ras. Critical to understanding the clinical effects of existing and future FTIs and geranylgeranyl transferase inhibitors (GGTIs), is the identification of in vivo substrates of FTase and GGTase-I. Our innovative strategy of using unnatural, transferable prenyl analogues and analogue specific monoclonal antibodies to identify prenylated cellular proteins will provide valuable information on the cellular targets of inhibitors of prenylation. We expect these studies to result in the identification of previously unknown prenylated proteins.
The specific aims of this project are: 1) Synthesis of farnesyl and geranylgeranyl diphosphate analogues to study protein prenylation. 2) Screening these compounds for ubstrate specificity of FTase and GGTase-I and development of prenyl function inhibitors. 3) Identification of prenylated proteins in cells. The results from these studies may provide leads to new molecules to treat cancer and also identify new molecular targets to develop anti-cancer therapies. Substantial evidence points to the central role of proteins normally modified by a prenyl group in cancer progression. We propose to design and test prenyl function inhibitors and to identify and characterize proteins that are modified with prenyl groups. The results from these studies may provide leads to new molecules to treat cancer and also identify new molecular targets to develop future anti-cancer therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM066152-07S1
Application #
8052002
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Fabian, Miles
Project Start
2010-05-15
Project End
2011-04-30
Budget Start
2010-05-15
Budget End
2011-04-30
Support Year
7
Fiscal Year
2010
Total Cost
$84,965
Indirect Cost
Name
University of Kentucky
Department
Biochemistry
Type
Schools of Medicine
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
Rush, Jeffrey S; Subramanian, Thangaiah; Subramanian, Karunai Leela et al. (2015) Novel Citronellyl-Based Photoprobes Designed to Identify ER Proteins Interacting with Dolichyl Phosphate in Yeast and Mammalian Cells. Curr Chem Biol 9:123-141
Rising, Kathleen A; Crenshaw, Charisse M; Koo, Hyun Jo et al. (2015) Formation of a Novel Macrocyclic Alkaloid from the Unnatural Farnesyl Diphosphate Analogue Anilinogeranyl Diphosphate by 5-Epi-Aristolochene Synthase. ACS Chem Biol 10:1729-36
Subramanian, Thangaiah; Ren, Hongmei; Subramanian, Karunai Leela et al. (2014) Design and synthesis of non-hydrolyzable homoisoprenoid ?-monofluorophosphonate inhibitors of PPAPDC family integral membrane lipid phosphatases. Bioorg Med Chem Lett 24:4414-4417
Degagné, Emilie; Pandurangan, Ashok; Bandhuvula, Padmavathi et al. (2014) Sphingosine-1-phosphate lyase downregulation promotes colon carcinogenesis through STAT3-activated microRNAs. J Clin Invest 124:5368-84
Srinivasan, Kamalakkannan; Subramanian, Thangaiah; Spielmann, H Peter et al. (2014) Identification of a farnesol analog as a Ras function inhibitor using both an in vivo Ras activation sensor and a phenotypic screening approach. Mol Cell Biochem 387:177-86
Chen, Min; Knifley, Teresa; Subramanian, Thangaiah et al. (2014) Use of synthetic isoprenoids to target protein prenylation and Rho GTPases in breast cancer invasion. PLoS One 9:e89892
Onono, Fredrick; Subramanian, Thangaiah; Sunkara, Manjula et al. (2013) Efficient use of exogenous isoprenols for protein isoprenylation by MDA-MB-231 cells is regulated independently of the mevalonate pathway. J Biol Chem 288:27444-55
Subramanian, Thangaiah; Subramanian, Karunai Leela; Sunkara, Manjula et al. (2013) Syntheses of deuterium labeled prenyldiphosphate and prenylcysteine analogues for in vivo mass spectrometric quantification. J Labelled Comp Radiopharm 56:370-5
Subramanian, Thangaiah; Pais, June E; Liu, Suxia et al. (2012) Farnesyl diphosphate analogues with aryl moieties are efficient alternate substrates for protein farnesyltransferase. Biochemistry 51:8307-19
Borowsky, Alexander D; Bandhuvula, Padmavathi; Kumar, Ashok et al. (2012) Sphingosine-1-phosphate lyase expression in embryonic and adult murine tissues. J Lipid Res 53:1920-31

Showing the most recent 10 out of 35 publications