Japan Science and Technology Corporation

Japan Science and Technology Agency

National Institute of Advanced Industrial Science and Technology

The University of Tokyo

Inter-University Research Institute Corporation National Institutes of Natural Sciences

Yoshio Umezawa

Moritoshi Sato

Takeaki Ozawa

Hiroshi Aoki

Hiroaki Tao

Akira Kanno

Takahito Ohshiro

SungBae Kim

Probefor detection and quantification of nitric oxide, which comprises two subunitsandof soluble guanylate cyclaseand cGMP-visualization probesrespectively connected with each subunit, wherein the cGMP-visualization probe generates signal upon recognizing guanosine 3',5&#x2032;-cyclic monophospate.

Probe for detection and quantification of nitric oxide, and method for detecting and quantifying nitric oxide using the same

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.

Activated protease indicator

A method is herein presented for analyzing nucleobases on a single molecular basis (a single molecule detection), which comprises scanning a molecular tip chemically modified with the complementary nucleobase on the nucleobases, and measuring the tunneling currents between the scanned nucleobases and the molecular tips with scanning tunneling microscopy.

Method for analyzing nucleobases on a single molecular basis

The claimed invention comprises a single molecule-format bioluminescent probe for detecting a target-specific ligand in a living cell, which comprises, a ligand-binding molecule of which conformation is changed upon binding to the ligand, wherein the ligand-binding molecule comprises a ligand-binding domain (LBD) of a nuclear receptor and an LBD-interacting domain that is a co-activator peptide of said nuclear receptor, and an N-terminal polypeptide and a C-terminal polypeptide of a click beetle luciferase (N-CBLuc and C-CBLuc), which flank each end of the ligand-binding molecule, respectively, wherein the N-CBLuc and the C-CBLuc self-complement to generate a luminescent signal only upon binding of the ligand to the ligand-binding molecule.

Single molecule-format bioluminescent probe

The present invention provides probes for detection and quantification of a lipid second messenger, which comprises: a polypeptide specifically bound to the lipid second messenger; two chromophores respectively having different fluorescence wavelengths, wherein each of the chromophores is linked to each end of the polypeptide through a rigid linker sequence; and a membrane localization sequence linked to one of the chromophores through a rigid linker sequence. According to the present invention, it is now possible to quantitatively detect when and in which site of a living cell the lipid second messengers are produced.

Probe for detecting and quantifying lipid second messenger and method of detecting and quantifying lipid second messenger using the same

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.

Probe for visualizing phosphorylation/dephosphorylation of protein and method of detecting and quantifying phosphorylation/dephosphorylation of protein

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.

Method of detection and quantification of cGMP by using a cGMP-visualizing probe

A convenient and commonly applicable method for the specific detection of a nucleic acid with an arbitrary sequence is provide. This method comprises attaching at least a nucleic acid single strand to an electrode, bringing the thus-obtained modified electrode in contact with a solution containing the analyte single-stranded nucleic acid, and measuring the redox reaction of the redox marker.

Electrochemical detection method of complementarity to nucleic acid bases

A probe for protein&#x2014;protein interaction analysis suitable for analyzing protein&#x2014;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&#x2014;protein interaction, and a physicochemically or biochemically detectable protein is regenerated.

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Tokyo, Japan

Yoshio Umezawa