Cells are able to process an enormous amount of infonnafion about their environment and their internal status via highly regulated signaling pathways. Critical to this information transfer are small molecule second messengers, such as cAMP, Ca^*, and a recently emerging family of phosphorylated inositides. Among the many derivatives of the inositol phosphate messengers is an intriguing subgroup that possesses high energy diphosphate groups, namely the diphosphoinositol phosphates (PP.-IPs), which have recently been connected to several cellular functions, including vesicular trafficking and apoptosis. The PP-IPs have also been linked to abnormal physiological processes including diminished insulin secretion and cancer. Owing to their chemically complex nature, there exist significant technical challenges to uncover inositol phosphate function using standard cell biology techniques. This proposal seeks to overcome these hurdles using chemical methods, to elucidate the discrete signaling functions of the PP-IPs. During the independent period of this award, aims '1,2, and 3 will rely on the deveiopment of chemical probes to study PP-IP function. Through the development of a reagent for the affinity purification of diphosphorylated proteins, the impact of this novel post-translational modification will be evaluated (aim 1). PP-IP analogues that can be covalently linked to their protein binding partners will serve to map the preferred inositol phosphate binding sites (aim 2). Lastly, aim 3 will explore the metal binding properties of the PP-IPs, which vwll then be exploited for the development of luminescent probes for in vivo imaging. Importantly, the tools from aims 1-3 m be evaluated in the cell models that have been generated during the mentored phase. The genetic interaction data will serve as a platform to generate and validate hypotheses about PP-IP signaling functions. The insulinoma cells are a critical model system to understand the role of PP-IPs in insulin secretion. By combining the chemical reagents with these genefic analyses, it will be possible to assess the physiological relevance of PP-IP signaling.

Public Health Relevance

The diphosphoinositol phosphates are an important class of secondary messengers in the cell, yet they have received little consideration to date. Their function has clearly been linked to several diseases, including the emerging diabetes epidemic. Elucidation of their signaling function will help to develop novel and improved therapeutics against the development to type 2 diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Transition Award (R00)
Project #
5R00GM087306-05
Application #
8325697
Study Section
Special Emphasis Panel (NSS)
Program Officer
Anderson, Vernon
Project Start
2009-04-01
Project End
2013-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
5
Fiscal Year
2012
Total Cost
$244,499
Indirect Cost
$78,409
Name
Princeton University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
002484665
City
Princeton
State
NJ
Country
United States
Zip Code
08544
Wu, Mingxuan; Chong, Lucy S; Capolicchio, Samanta et al. (2014) Elucidating diphosphoinositol polyphosphate function with nonhydrolyzable analogues. Angew Chem Int Ed Engl 53:7192-7
Marmelstein, Alan M; Yates, Lisa M; Conway, John H et al. (2014) Chemical pyrophosphorylation of functionally diverse peptides. J Am Chem Soc 136:108-11
Wu, Mingxuan; Dul, Barbara E; Trevisan, Alexandra J et al. (2013) Synthesis and characterization of non-hydrolysable diphosphoinositol polyphosphate second messengers. Chem Sci 4:405-410