35 results about "Kinase substrate" patented technology

Disclosed are fluorescent compositions and methods for detecting and / or characterizing enzymes and various uses thereof.

The present invention relates to apparatus and methods for separating and detecting enzyme substrates using separation gels. For example, the apparatus and methods can be used to separate and detect kinase substrates for further analysis. The apparatus and methods can also be used to detect enzyme inhibitors, such as kinase inhibitors.

An in vitro protein kinase assay technology that (1) exhibits a high assay signal to background ratio (S / B) and range (S-B); (2) is homogenous; (3) is non-radioactive; and (4) does not require a phospho-specific antibody involves complexing a trivalent metal ion (e.g. Ga<3+>, Fe<3+>, Al<3+>, In<3+>, Ru<3+>, Sc<3+>, Y<3+>) to the surface of amplified luminescent proximity assay acceptor or donor beads, e.g., via a suitable linker such as nitrilotriacetic acid (NTA; also referred to as carboxymethyl-lysine), iminodiacetic acid (IDA), or an appropriately substituted N-containing heterocycle, for example a triazoheterocycle, for example a triazocyclononaneononane, such as 1-propylamino-4-acetato-1,4,7-triazacyclononane. A protein (or constituent part) or other kinase substrate is bound to the surface of the other of an amplified luminescent proximity assay acceptor or donor bead and, if phosphorylated, brought into proximity with the trivalent metal ion-complexed acceptor bead to generate a luminescent signal. Presence of a kinase inhibitor inhibits phosphorylation and therefore signal generation and, in this way, is detectable. As the invention described herein recognizes the presence or absence of phosphate groups on a protein, (or constituent part), or other biological macromolecule (e.g., mono, di, or trinucleotides, cyclic nucleotides or phosphate substituted inositols), it is broadly applicable to any phosphorlylation or dephosphorylation reaction enzymes and provides a highly robust and flexible assay format for protein kinases and other enzyme classes, including lipid kinases, phosphatases, phosphodiesterases and others.

The invention provides kinase substrates and methods comprising their use.

The invention discloses the application of hepatocyte growth regulator tyrosine kinase substrate (HGS) in preventing and / or treating EV71 infection, and belongs to the field of antiviral drugs. The present invention detects the mRNA level and protein level of EV71 VP1 by adding HGS protein into the cells and adding EV71, and finds that the mRNA and protein levels of VP1 decrease significantly compared with those without adding HGS, and it is in a dose-dependent manner, indicating that HGS has an anti-inflammatory effect. The effect of EV71. The effect of HGS on the survival rate of EV71-infected cells was further detected, and it was found that the survival rate of cells added with HGS protein was significantly higher than that of the virus control group. Preventive and therapeutic effects. The present invention discovers the new medicinal value of HGS, which can be used for preparing anti-EV71 medicine, medicine for preventing and / or treating EV71 infection.

The present invention relates to a biochip for detecting phosphorylation and a method for detecting phosphorylation using the same, more precisely a biochip prepared by integrating a protein produced from the recombination of a substrate of kinase selected from the group consisting of PKC (Protein Kinase C), cdc2-PK (cdc2 Protein Kinase) and DNA-PK (DNA-dependent Protein Kinase) and the elevated protein such as Selenomonas ruminantium membrane protein on a matrix surface coated with an active group, a kit for detecting phosphorylation composed of the said biochip and a cofactor labeled with a radio-isotope and a method for detecting phosphorylation using the same. The biochip for detecting phosphorylation of the present invention using a radio-isotope facilitates the detection of phosphorylation with a minimum amount of a sample by simple processes, compared with the conventional method using an antibody. Since this method can analyze a large amount of samples in a shorter period of time, it can be effectively used for the analysis of kinase activity.

The invention provides an LRR (leucine-rich repeat) receptor kinase substrate searching method. The LRR receptor kinase substrate searching method includes: S1, incubating LRR receptor kinase along with ATP-gamma-S to obtain a first mixture; S2, incubating the first mixture along with total protein which is rich in LRR receptor kinase structures so as to obtain a second mixture; S3, incubating thesecond mixture along with PNBM to obtain a third mixture; S4, removing the PNBM in the third mixture to obtain a fourth mixture; S5, incubating the fourth mixture with pretreated protein G agarose toobtain first supernatant; S6, incubating the first supernatant along with a phosphorothioate resistant monoclonal antibody, which is in coupled linkage with the protein G agarose, so as to obtain first sediment; S7, analyzing the first sediment to determine an LRR receptor kinase substrate.

The invention discloses a mycobacterium M. tuberculosis holoprotein chip and an application thereof. The protein chip has at least one of the following functions (1) to (7); the following 4,168 proteins are connected to the protein chip, and each protein independently forms a detection point. Experiments show that the mycobacterium M. tuberculosis holoprotein chip consists of a substrate and proteins containing more than 95 percent of mycobacterium M. tuberculosis genome codes. The holoprotein chip is obviously significant for looking for a serum biomarker, a substrate interacting with a micromolecular compound c-di-GMP, a kinase substrate interacting with protein kinase and a substrate interacting with macrophage. Compared with an existing research means, the holoprotein chip has global and high-throughput screening characteristics, research design and operation processes are simplified, and the research efficiency is remarkably improved.