Chimeric phosphorylation indicator
US8669074B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 28, 2004 |
| Grant date | Mar 11, 2014 |
| Priority date | — |
| Expiry date | Nov 10, 2031 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/43595
- WIPO fieldMeasurement
- WIPO sectorInstruments
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.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.