Assays for protein kinases using fluorescent protein substrates
First Claim
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1. A polynucleotide, comprisinga nucleic acid molecule encoding an Aequorea-related fluorescent protein comprising a phosphorylation site for a protein kinase within ten amino acids of the amino terminus of said Aequorea-related fluorescent protein,wherein said Aequorea-related fluorescent protein exhibits a different fluorescent property in the phosphorylated state than in the un-phosphorylated state.
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Abstract
This invention provides assays for protein kinase activity using fluorescent proteins engineered to include sequences that can be phosphorylated by protein kinases. The proteins exhibit different fluorescent properties in the non-phosphorylated and phosphorylated states.
171 Citations
23 Claims
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1. A polynucleotide, comprising
a nucleic acid molecule encoding an Aequorea-related fluorescent protein comprising a phosphorylation site for a protein kinase within ten amino acids of the amino terminus of said Aequorea-related fluorescent protein, wherein said Aequorea-related fluorescent protein exhibits a different fluorescent property in the phosphorylated state than in the un-phosphorylated state.
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16. A polynucleotide, comprising:
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a nucleic acid encoding an Aequorea-related fluorescent protein with at least a contiguous sequence of 200 amino acids having at least 95% identity with the amino acid sequence of SEQ ID NO;
2,wherein said Aequorea-related fluorescent protein is modified within ten amino acids of its amino-terminus by insertion of a phosphorylation site for a protein kinase, and further wherein said Aequorea-related fluorescent protein exhibits a different fluorescent property in the phosphorylated state than in the unphosphorylated state. - View Dependent Claims (17, 18, 19, 20, 21, 22)
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23. A host cell, comprising:
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a recombinant nucleic acid molecule, said recombinant nucleic acid molecule comprising expression control sequences operatively linked to a nucleic acid sequence encoding an Aequorea-related fluorescent protein, wherein said Aequorea-related fluorescent protein comprises a phosphorylation site for a protein kinase within ten amino acids of its amino terminus, wherein said Aequorea-related fluorescent protein exhibits a different fluorescent property in the phosphorylated state than in the unphosphorylated state.
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Specification