Abstract: The present invention intends to provide an assay system using split luciferase that has a remarkably high detection sensitivity. In an embodiment, binding of mutually binding first and second proteins is detected by preparing a first fusion protein comprising the first protein fused with a peptide having the amino acid sequence of amino acid SEQ ID NO: 1 and a second fusion protein comprising the second protein fused with a peptide having an amino acid sequence selected from the group consisting of amino acid SEQ ID NOS: 2 to 6, and allowing the first fusion protein to bind with the second fusion protein to form a complex, and detecting luminescence emitted from the complex.

Abstract: The present invention intends to provide an assay system using split luciferase that has a remarkably high detection sensitivity. In an embodiment, binding of mutually binding first and second proteins is detected by preparing a first fusion protein comprising the first protein fused with a peptide having the amino acid sequence of amino acid SEQ ID NO: 1 and a second fusion protein comprising the second protein fused with a peptide having an amino acid sequence selected from the group consisting of amino acid SEQ ID NOS: 2 to 6, and allowing the first fusion protein to bind with the second fusion protein to form a complex, and detecting luminescence emitted from the complex.

Abstract: It is an object of the invention to provide novel approaches capable of detecting activated protease and also detecting protease activation on real time at a high sensitivity in a noninvasive manner. By the method for detecting activated protease of the invention, an indicator in a circular form comprising the C-half fragment of luciferase (Luc-C) and the N-half fragment of luciferase (Luc-N) linked together through a substrate peptide for a protease is introduced in an in vitro assay system or in cells. Upon digestion of the substrate peptide by the protease, Luc-N and Luc-C together reconstructs active luciferase, so that the activated protease can be detected by assaying the luminescence signal from the luciferase.

Abstract: The present invention intends to provide an assay system using split luciferase that has a remarkably high detection sensitivity. In an embodiment, binding of mutually binding first and second proteins is detected by preparing a first fusion protein comprising the first protein fused with a peptide having the amino acid sequence of amino acid SEQ ID NO: 1 and a second fusion protein comprising the second protein fused with a peptide having an amino acid sequence selected from the group consisting of amino acid SEQ ID NOS: 2 to 6, and allowing the first fusion protein to bind with the second fusion protein to form a complex, and detecting luminescence emitted from the complex.

Abstract: The present invention intends to provide an assay system using split luciferase that has a remarkably high detection sensitivity. In an embodiment, binding of mutually binding first and second proteins is detected by preparing a first fusion protein comprising the first protein fused with a peptide having the amino acid sequence of amino acid SEQ ID NO: 1 and a second fusion protein comprising the second protein fused with a peptide having an amino acid sequence selected from the group consisting of amino acid SEQ ID NOS: 2 to 6, and allowing the first fusion protein to bind with the second fusion protein to form a complex, and detecting luminescence emitted from the complex.

Abstract: It is an object of the invention to provide novel approaches capable of detecting activated protease and also detecting protease activation on real time at a high sensitivity in a noninvasive manner. By the method for detecting activated protease of the invention, an indicator in a circular form comprising the C-half fragment of luciferase (Luc-C) and the N-half fragment of luciferase (Luc-N) linked together through a substrate peptide for a protease is introduced in an in vitro assay system or in cells. Upon digestion of the substrate peptide by the protease, Luc-N and Luc-C together reconstructs active luciferase, so that the activated protease can be detected by assaying the luminescence signal from the luciferase.

Abstract: A convenient and highly accurate method for detecting protein nuclear transport induced by an endogenous or exogenous substance in local areas of living cells or animals is provided. The method uses a pair of probes for detecting protein nuclear transport, comprising Probe I and Probe II. In Probe I, a protein whose nuclear transport is to be detected or quantified is connected to an N-terminal end or a C-terminal end of a fusion protein [intein-C/reporter protein-C] wherein at least a C-terminal side polypeptide of an intein and a C-terminal side polypeptide of a reporter protein are connected in this order, and in Probe II, a nuclear localization signal is connected to an N-terminal end or a C-terminal end of a fusion protein [reporter protein-N/intein-N] wherein at least the remaining N-terminal side polypeptide of the reporter protein and the remaining N-terminal side polypeptide of the intein are connected in this order.

Abstract: A method for analyzing an organelle-localized protein, which enables one to determine whether or not a test protein localizes to an organelle, comprising the steps of: (a) a step of introducing a fusion peptide (a), which comprises one half-peptide of an intein, one half-peptide of a fluorescent protein and an organelle-targeting signal peptide, into a eukaryotic cell; (b) a step of introducing a test protein bound to a fusion peptide (b), which comprises the other half-peptide of the fluorescent protein and the other half-peptide of the intein, into the eukaryotic cell; and (c) a step of detecting fluorescence signal emitted by the fluorescent protein, and a material for analysis to be used in such method are provided.

Abstract: As a versatile method of detecting and assaying intracellular protein phosphorylation and dephosphorylation that enables nondestructive monitoring as well as spatial and temporal analysis for living cells, animal bodies, plant bodies and the like, a probe for imaging protein phosphorylation and dephosphorylation, which comprises a tandem fusion unit composed of a substrate domain that contains a phosphorylation and dephosphorylation site, a linker sequence and a phosphorylation recognition domain, interposed between a donor chromophore and an acceptor chromophore that cause fluorescence resonance energy transfer, is used.

Abstract: As a cGMP-visualizing probe capable of detecting and quantifying cGMP easily and accurately even in vivo and for a method of detecting and quantifying cGMP by using the same, a cGMP-visualizing probe comprising a polypeptide, which binds specifically to cGMP, and two chromophores having different fluorescence wavelengths each linked respectively to the two terminals of said polypeptide is provided.

Abstract: A convenient and highly accurate method for detecting protein nuclear transport induced by an endogenous or exogenous substance in local areas of living cells or animals is provided. The method uses a pair of probes for detecting protein nuclear transport, comprising Probe I and Probe II. In Probe I, a protein whose nuclear transport is to be detected or quantified is connected to an N-terminal end or a C-terminal end of a fusion protein [intein-C/reporter protein-C] wherein at least a C-terminal side polypeptide of an intein and a C-terminal side polypeptide of a reporter protein are connected in this order, and in Probe II, a nuclear localization signal is connected to an N-terminal end or a C-terminal end of a fusion protein [reporter protein-N/intein-N] wherein at least the remaining N-terminal side polypeptide of the reporter protein and the remaining N-terminal side polypeptide of the intein are connected in this order.

Abstract: The present invention provides a pair of probes for analyzing protein-protein interactions, which comprises a probe A containing at least an N-terminal half polypeptide of split Renilla luciferase, and a probe B containing at least the remaining C-terminal half polypeptide of split Renilla luciferase.

Abstract: A probe for protein—protein interaction analysis suitable for analyzing protein—protein interactions of various proteins with high accuracy and in a simple manner and a method for analyzing interaction of two proteins by using the probe. With the probe, protein splicing is caused by protein—protein interaction, and a physicochemically or biochemically detectable protein is regenerated.

Abstract: As a cGMP-visualizing probe capable of detecting and quantifying cGMP easily and accurately even in vivo and for a method of detecting and quantifying cGMP by using the same, a cGMP-visualizing probe comprising a polypeptide, which binds specifically to cGMP, and two chromophores having different fluorescence wavelengths each linked respectively to the two terminals of said polypeptide is provided.

Abstract: As a versatile method of detecting and assaying intracellular protein phosphorylation and dephosphorylation that enables nondestructive monitoring as well as spatial and temporal analysis for living cells, animal bodies, plant bodies and the like, a probe for imaging protein phosphorylation and dephosphorylation, which comprises a tandem fusion unit composed of a substrate domain that contains a phosphorylation and dephosphorylation site, a linker sequence and a phosphorylation recognition domain, interposed between a donor chromophore and an acceptor chromophore that cause fluorescence resonance energy transfer, is used.

Abstract: As a cGMP-visualizing probe capable of detecting and quantifying cGMP easily and accurately even in vivo and for a method of detecting and quantifying cGMP by using the same, a cGMP-visualizing probe comprising a polypeptide, which binds specifically to cGMP, and two chromophores having different fluorescence wavelengths each linked respectively to the two terminals of said polypeptide is provided.

Abstract: A method for analyzing an organelle-localized protein, which enables one to determine whether or not a test protein localizes to an organelle, comprising the steps of: (a) a step of introducing a fusion peptide (a), which comprises one half-peptide of an intein, one half-peptide of a fluorescent protein and an organelle-targeting signal peptide, into a eukaryotic cell; (b) a step of introducing a test protein bound to a fusion peptide (b), which comprises the other half-peptide of the fluorescent protein and the other half-peptide of the intein, into the eukaryotic cell; and (c) a step of detecting fluorescence signal emitted by the fluorescent protein, and a material for analysis to be used in such method are provided.

Abstract: As a versatile method of detecting and assaying intracellular protein phosphorylation and dephosphorylation that enables nondestructive monitoring as well as spatial and temporal analysis for living cells, animal bodies, plant bodies and the like, a probe for imaging protein phosphorylation and dephosphorylation, which comprises a tandem fusion unit composed of a substrate domain that contains a phosphorylation and dephosphorylation site, a linker sequence and a phosphorylation recognition domain, interposed between a donor chromophore and an acceptor chromophore that cause fluorescence resonance energy transfer, is used.

Abstract: A probe for protein-protein interaction analysis applicable to various proteins, which enables simple analysis in high accuracy, and a method for analyzing interaction of two proteins is provided. Protein splicing from the probe is caused by protein-protein interaction, and a physiochemically or biochemically detectable protein is regenerated.

Abstract: As a cGMP-visualizing probe capable of detecting and quantifying cGMP easily and accurately even in vivo and for a method of detecting and quantifying cGMP by using the same, a cGMP-visualizing probe comprising a polypeptide, which binds specifically to cGMP, and two chromophores having different fluorescence wavelengths each linked respectively to the two terminals of said polypeptide is provided.