Engineering a phosphotyrosyl-tRNA Synthetase

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R41GM113301-01
Agency Tracking Number: R41GM113301
Amount: $220,323.00
Phase: Phase I
Program: STTR
Solicitation Topic Code: 300
Solicitation Number: PA14-072
Timeline
Solicitation Year: 2015
Award Year: 2015
Award Start Date (Proposal Award Date): 2015-02-01
Award End Date (Contract End Date): 2016-08-31
Small Business Information
688 E MAIN ST, STE 2A, Branford, CT, 06405-2971
DUNS: 078402317
HUBZone Owned: N
Woman Owned: N
Socially and Economically Disadvantaged: N
Principal Investigator
 MICHAEL WEINER
 (203) 458-2844
 michael@mweiner.com
Business Contact
 MICHAEL WEINER
Phone: (203) 606-5394
Email: mweiner@axiomxinc.com
Research Institution
 COPPIN STATE UNIVERSITY
 2500 W. NORTH AVENUE
BALTIMORE, MD, 21216-3698
 Nonprofit college or university
Abstract
DESCRIPTION provided by applicant Although tyrosine phosphorylation is an important controlling element in cell signaling no tRNA suppressor for phosphotyrosine pTyr incorporation has yet been made We propose to use directed molecular evolution of several aminoacyl tRNA synthetases aaRSs to identify mutations that enable binding of pTyr to the aaRS Specifically ATP molecules will be attached to beads which will then be incubated with free pTyr and a phage display library of a mutated aaRS If a mutant aaRS can catalyze the formation of pTyr AMP which is the intermediate for the generation of charged tRNA it will bind to the beads and can be enriched The mutated aaRSs will be used in in vitro translation to incorporate the pTyr into the protein structure of an assayable gene for example galactosidase Mass Spectrometry and a set of already existing anti pTyr specific antibodies will be used to validate incorporation of the pTyr in the assayable protein Phase I is focused on in vitro incorporation PUBLIC HEALTH RELEVANCE The ability to generate tyrosyl phosphorylated proteins will have significant utility in studying the role of phosphoproteins that are involved in cell signalin inflammation cancer and other diseases Conventional approaches toward making phosphorylated proteins require kinases which are promiscuous and often lead to phosphorylation at multiple undesirable residues Successful completion of this proposal will be the first method to allow site specific incorporation of phosphotyrosine in a protein

* Information listed above is at the time of submission. *

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