Phosphohistidine Mimetics for the Development of Anti-Phosphohistidine Antibodies and Identification of Therapeutic Targets

Web Published:
1/26/2015
Description:

Phosphohistidine Mimetics for the Development of Anti-Phosphohistidine Antibodies and Identification of Therapeutic Targets

Princeton Docket # 14-2972

Researchers in the Department of Chemistry at Princeton University have developed the first phosphohistidine (pHis)-specific antibody, which has enabled the selective detection and identification of pHis proteins from complex biological samples.  Protein phosphorylation plays a critical role in central metabolism and cell signaling and is one of the most common posttranslational modifications.  Misregulation of protein phosphorylation can lead to disease states.  Histidine is one amino acid that can be phosphorylated, and histidine kinase activity is upregulated in cancer cells.

 

Studies on pHis have been hampered by its chemical instability.  The development of stable pHis analogues will advance antibody production and studies of histidine phosphorylation.  The Muir lab has synthesized a second-generation pHis analog where the triazole heterocycle in the first generation mimic has been replaced with a pyrazole.  These pyrazole-based pHis analogs will be used for the development of new pHis antibodies and the identification of therapeutic targets.

 

Applications:   

·         Identify drug targets in protein kinases and phosphatases

·         Develop haptens and immunogens for antibody production and conjugates

·         pHis antibodies used as inhibitors against protein phosphohistidine phosphatases

·         Study protein phosphorylation and misregulation leading to disease

·         Identify physiological functions of pHis in cell signaling, metabolism, and epigenetics

Advantages: 

·         Phosphohistidine (pHis) analog incorporation into peptides and proteins

·         Hydrolytically stable against acid or phosphatases

·         Improved molecular design and mimic of pHis with pyrazole-based analog

·         Easily synthesized from commercial and previously known materials

·         High specificity and potency

·         Detection of phosphorylated histidine in biological samples

Keywords

Protein phosphorylation, posttranslational modification, phosphohistidine, antibody, therapeutic target

 

Related Publications

Kee, J.-M., Oslund, R.C., Couvillon, A.D., and Muir, T.W.  A second-generation phosphohistidine analog for production of phosphohistidine antibodies.  Organic Letters.  2014, in press.

 

Oslund R.C., Kee, J.-M., Couvillon, A.D., Perlman, D.H., and Muir, T.W.  A phosphohistidine proteomics strategy based on elucidation of a unique gas-phase phosphopeptide fragmentation mechanism.  Journal of the American Chemical Society.  2014, 136, 12899-12911.

 

Kee, J.-M., Oslund, R.C., Perlman, D.H., and Muir, T.W.  A pan-specific antibody for direct detection of protein histidine phosphorylation.  Nature Chemical Biology.  2013, 9, 416-421.

 

Kee, J.-M., Villani, B., Carpenter, L.R., and Muir, T.W.  Development of Stable Phosphohistidine Analogues.  Journal of the American Chemical Society.  2010, 132, 14327-14329.

 

Kee, J.-M., and Muir, T.W.  Phosphohistidine analogs (PCT/US2011/052854, WO2012040523 A3), The Rockefeller University.

 

Inventors

 

Rob C. Oslund, Ph.D., Jung-Min Lee, Ph.D., and Tom W. Muir, Ph.D.

 

Faculty inventor

 

Tom W. Muir, Van Zandt Williams, Jr. Class of ’65 Professor of Chemistry and Department Chair

 

The Muir lab investigates the physiochemical basis of protein function in complex systems of biomedical interest.  By combining tools of synthetic chemistry, protein biochemistry, and cell biology, the Muir lab has developed a suite of new technologies that provide fundamental insight into how proteins work, including the intein splicing reaction and protein ligation as a platform for chemical biology studies.

 

In 2011, Dr. Muir joined the Princeton University faculty as the Van Zandt Williams Jr. Class of ’65 Professor of Chemistry. In 2015, he became the Chair of the Department of Chemistry. He has published over 150 scientific articles and has won a number of honors for his research, including the Burrough Wellcome Fund New Investigator Award, the Pew Award in the Biomedical Sciences, the Alfred P. Sloan Research Fellow Award, the Leonidas Zervas Award from the European Peptide Society, the Irving Sigal Award from the Protein Society, the 2008 Vincent du Vigneaud Award in Peptide Chemistry, the 2008 Blavatnik Award from the New York Academy of Sciences, the 2008 Distinguished Teaching Award from The Rockefeller University, the 2012 Jeremy Knowles Award from the Royal Society of Chemistry, and a 2013 Arthur C. Cope Scholar Award from the American Chemical Society.  Prof. Muir is the recipient of a MERIT Award from the US National Institutes of Health and is a Fellow of American Association for the Advancement of Science and the Royal Society of Edinburgh.

 

Prof. Muir received his B.Sc (Hons, 1st class) in Chemistry from the University of Edinburgh in 1989 and his Ph.D. in Chemistry from the same institute in 1993 under the direction of Professor Robert Ramage. After postdoctoral studies with Stephen B.H. Kent at the Scripps Research Institute, he joined the faculty at the Rockefeller University in New York City in 1996, where he was, until recently, the Richard E. Salomon Family Professor and Director of the Pels Center of Chemistry, Biochemistry and Structural Biology.

 

Intellectual Property and Licensing Status

Patent applications are pending.  Princeton is seeking industrial collaborators for further development and commercialization of this technology.

 

Contact

Laurie Tzodikov
Princeton University Office of Technology Licensing • (609) 258-7256•
tzodikov@princeton.edu

 

Sarah Johnson

Princeton University Office of Technology Licensing • sajohnso@princeton.edu

 

 

 

Patent Information:
For Information, Contact:
Prabhpreet Gill
Licensing Associate
Princeton University
psgill@princeton.edu
Inventors:
Tom Muir
Rob Oslund
Jung-Min Kee
Keywords:
antibody
biomarker/diagnostic
Biotechnology/Pharmaceuticals
Chemistry
drug discovery
drug target
life science research tools
posttranslational modification
protein engineering
protein phosphorylation
therapeutic