Invention Grant
- Patent Title: Chimeric phosphorylation indicator
- Patent Title (中): 嵌合磷酸化指示剂
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Application No.: US10857622Application Date: 2004-05-28
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Publication No.: US08669074B2Publication Date: 2014-03-11
- Inventor: Jonathan D. Violin , Alexandra C. Newton , Roger Y. Tsien , Jin Zhang
- Applicant: Jonathan D. Violin , Alexandra C. Newton , Roger Y. Tsien , Jin Zhang
- Applicant Address: US CA Oakland
- Assignee: The Regents of the University of California
- Current Assignee: The Regents of the University of California
- Current Assignee Address: US CA Oakland
- Agency: Morgan, Lewis & Bockius LLP
- Main IPC: C12P21/04
- IPC: C12P21/04

Abstract:
A chimeric phosphorylation indicator (CPI) as provided herein can contain a donor molecule, a phosphorylatable domain, a phosphoaminoacid binding domain (PAABD), and an acceptor molecule. Where the phosphorylatable domain is phosphorylatable by protein kinase C (PKC), the CPI is a c-kinase activity reporter (CKAR). Donor and acceptor molecules may be, independently, fluorescent proteins such as non-oligomerizing fluorescent proteins. A CPI can contain a phosphorylatable polypeptide and a fluorescent protein; the phosphorylatable polypeptide may be contained within the sequence of the fluorescent protein, or the fluorescent protein may be contained within the sequence of the phosphorylatable polypeptide. The spatiotemporal properties of the PKC signal pathway may be tested with CKAR, calcium-sensing fluorophores and FRET-based translocation assays. Polynucleotides encoding such CPIs, and kits containing the indicators and/or the polynucleotides, are provided. A method of using the chimeric phosphorylation indicators to detect a kinase or phosphatase in a sample is provided.
Public/Granted literature
- US20050026234A1 Emission ratiometric indicators of phosphorylation by C-kinase Public/Granted day:2005-02-03
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