Abstract: To provide a semaphorin inhibitor; a peripheral or central nerve regeneration promoter which contains said semaphorin inhibitor as an active ingredient; and a preventive or remedy for a neuropathic disease and a neurodegenerative disease containing said nerve regeneration promoter, or the like. A low-molecular weight compound, which acts at a concentration of 10 ?g/ml or below to inhibit the growth cone collapse activity of semaphorin such as semaphorin 3A, semaphorin 6C or the like and/or the nerve outgrowth inhibitory activity of semaphorin in a collagen gel and which does not substantially affect cell proliferation, is obtained from the culture of strain SPF-3059 belonging to the genus Penicillium. The low-molecular weight compound with the semaphorin inhibitory activity thus obtained exhibits the in vivo nerve-regeneration promoting action.

Abstract: To provide a semaphorin inhibitor; a peripheral or central nerve regeneration promoter which contains said semaphorin inhibitor as an active ingredient; and a preventive or remedy for a neuropathic disease and a neurodegenerative disease containing said nerve regeneration promoter, or the like. A low-molecular weight compound, which acts at a concentration of 10 ?g/ml or below to inhibit the growth cone collapse activity of semaphorin such as semaphorin 3A, semaphorin 6C or the like and/or the nerve outgrowth inhibitory activity of semaphorin in a collagen gel and which does not substantially affect cell proliferation, is obtained from the culture of strain SPF-3059 belonging to the genus Penicillium. The low-molecular weight compound with the semaphorin inhibitory activity thus obtained exhibits the in vivo nerve-regeneration promoting action.

Abstract: This invention has as its object a method for detecting catalytic activity of a sample, characterized in that it comprises: the incubation of a substrate (S) with the sample that may have the catalytic activity that it is desired to detect, the addition of a reagent (X) that can react either with a chemical group of unconsumed substrate (S) or with a chemical group of product (P) that is formed after an incubation period with the sample, the addition of a developer (R) that can react with reagent (X), and the detection of the transformation of developer (R).

Abstract: Method for measuring the presence or absence of chemical groups, in particular phosphate groups, attached to biological molecules in a sample in which these molecules are tagged with fluorescent markers and these fluorescent markers are activated by means of irradiating the sample with light. The method is characterized by the following steps: a) Use of a fluorescent marker, the fluorescence lifetime of which assumes a different value depending upon the presence or absence of phosphate groups attached to the biomolecule; b) Measurement of the fluorescence lifetime of the fluorescent marker attached to a biomolecule and selected in accordance with Step a); c) Classification of the biomolecules in accordance with the presence or absence of phosphate groups attached to these, based on the different lifetime of each.

Abstract: Described herein are methods which identify candidate agents as binding to a protein or as a modulator of the binding characteristics or biological activity of a protein. Generally, the methods involve the use of ADP or phosphate. The assays can be used in a high throughput system to obviate the cumbersome steps of using gels or radioactive materials.

Abstract: A method is disclosed whereby the concentration of a blood substitute, such as cross-linked hemoglobin, in a serum or plasma specimen is rapidly and accurately identified and quantified. The method further takes the measured concentration of the blood substitute and uses it to correct for its effect, if any, on a measured analyte concentration, e.g., serum/plasma total protein. Further, the method allows for the determination of the concentration of true hemoglobin in the presence of blood substitutes. The method is carried out in respect of samples contained in a primary or secondary labelled tube, or a pipette tip used to dispense serum or plasma in a blood analyzer.

Abstract: PAK4 and JIK, both of which bind to MKK7 and directly phosphorylate MKK7, were found in the present invention. The present invention provides an inhibitor of c-Jun phosphorylation caused by JNK3 and a method for inhibiting the same, and an agent for preventing and/or treating a disorder attributable to c-Jun phosphorylation caused by JNK3 and a method for preventing and/or treating the same, all of which comprise inhibiting one member selected from the following: the binding of PAK4 to MKK7, the phosphorylation of MKK7 by PAK4, the binding of JIK to MKK7, and the phosphorylation of MKK7 by JIK. Further, the present invention provides a method for identifying a compound that inhibits the binding of PAK4 to MKK7, the phosphorylation of MKK7 caused by PAK4, the binding of JIK to MKK7, or the phosphorylation of MKK7 caused by JIK, as well as the compound obtained thereby.

Abstract: The present invention teaches a marker useful for detection and measurement of free radical damage. Specifically, the invention takes advantage of alterations which occur to the N-terminus of the albumin molecule, a circulating protein in human blood, in the presence of free radicals. These alterations effect the ability of the N-terminus of the albumin molecule to bind metals. Methods for detecting and quantifying this alteration include evaluating and quantifying the cobalt binding capacity of an albumin-containing sample, analysis and measurement of the ability of albumin to bind exogenous cobalt, detection and measurement of the presence of copper in a purified albumin sample and use of an immunological assay specific to the altered form of serum albumin which occurs following free radical damage.

Abstract: A novel gene defining a novel human UDP-GlcNAc: Gal/Gl cNAc? 1-3GalNAc ??1, 6GlcNAc-transferase, termed C2/4GnT, with unique enzymatic properties is disclosed. The enzymatic activity of C2/4GnT is shown to be distinct from that of previously identified enzymes of this gene family. The invention discloses isolated DNA molecules and DNA constructs encoding C2/4GnT and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting C2/4GnT activity, as well as cloning and expression vectors including such DNA, cells transfected with the vectors, and recombinant methods for providing C2/4GnT. The enzyme C2/4GnT and C2/4GnT-active derivatives thereof are disclosed, in particular soluble derivatives comprising the catalytically active domain of C2/4GnT.

Abstract: Compounds that inhibit ubiquitin-mediated proteolysis of phosphorylated I?B by interfering, directly or indirectly, with the ability of ?-TrCP/E3RS to engage in protein-protein association involving hnRNP-U, are useful as drugs for treating conditions associated with NF-?B activation. Cellular and non-cellular screening methods for identifying such compounds are based on monitoring the association/dissociation of ?-TrCP/E3RS.

Abstract: The invention relates to isolated nucleotide sequences from Coryneform bacteria which code for the pck gene encoding the enzyme phosphoenol pyruvate carboxykinase (PEP carboxykinase). The invention also relates a process for the fermentative preparation of L-amino acids, in particular L-lysine, L-threonine, and L-glutamate by attenuation of the pck gene.

Abstract: Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. A method to detect proteins which contain nitrosothiols involves several steps. Nitrosylated cysteines are converted to tagged cysteines. Tagged proteins can then be detected, for example, by immunoblotting and/or can be purified by affinity chromatography. The method is applicable to the detection of S-nitrosylated proteins in cell lysates following in vitro S-nitrosylation, as well as to the detection of endogenous S-nitrosothiols in selected protein substrates.

Abstract: Methods for rapidly identifying drug candidates that can bind to an enzyme at both a common ligand site and a specificity ligand site, resulting in high affinity binding. The bi-ligand drug candidates are screened from a focused combinatorial library where the specific points of variation on a core structure are optimized. The optimal points of variation are identified by which atoms of a ligand bound to the common ligand site are identified to be proximal to the specificity ligand site. As a result, the atoms proximal to the specificity ligand site can then be used as a point for variation to generate a focused combinatorial library of high affinity drug candidates that can bind to both the common ligand site and the specificity ligand site. Different candidates in the library can then have high affinity for many related enzymes sharing a similar common ligand site.

Abstract: Apparatus and method for determining concentration of creatinine in whole blood or plasma using a 1-methylhydantoinase (NMHase) catalyzed reaction, wherein the (NMHase) used in the reaction has its own substrate, NMH, bound thereto. The present invention uses commercially available NMHase and thus eliminates prior art stabilization procedures for NMHase. Conveniently, known Trinder reagents and oxidative couplers are used for the indicator system. It has been found that by judiciously selecting the reagents used for the indicator system and/or varying the amount of enzyme NMHase that is loaded into the assay, the effect of the blank reaction can be minimized in the dynamic range of interest such that concentration of creatinine in normal and pathological levels can be measured directly without having to adjust for a blank reaction caused by bound NMH.

Abstract: Amounts of components in a specimen can be analyzed with excellent quantitativity. The analysis includes: measuring an amount of a component to be analyzed in a specimen; measuring an amount of a standard component present originally and homeostatically in the specimen other than the component to be analyzed; determining the amount of the specimen from the amount of the standard component thus measured and a known concentration of the standard component in the specimen; and determining a concentration of the component to be analyzed in the specimen from the amount of the specimen thus determined and the amount of the component to be analyzed thus measured. The quantitative analysis of the present invention allows a component to be analyzed to be measured with high quantitativity as shown in FIG. 1.

Abstract: Described herein are methods which identify candidate agents as binding to a protein or as a modulator of the binding characteristics or biological activity of a protein. Generally, the methods involve the use of ADP or phosphate. The assays can be used in a high throughput system to obviate the cumbersome steps of using gels or radioactive materials.

Abstract: The invention provides isolated nucleic acid and amino acid sequences of HsKifC2, antibodies to HsKifC2, methods of screening for HsKifC2 modulators using biologically active HsKifC2, and kits for screening for HsKifC2 modulators.

Abstract: A method for measuring stains of a test sample by measuring adenosine phosphates. Luminescence of the adenosine phosphates is induced by a luminescent reagent (33) containing an ATP regenerating enzyme. The level or quantity of luminescence is detected by a silicon photodiode (5) to thereby measure the stains.

Abstract: Substituted 4-hydroxy-2-furoic acids are obtained by the fermentation of an asterriquinone, which is a natural product having a 2,5-dioxy-3,6-bis(indolyl)quinone structure. The compounds modulate insulin receptor tyrosine kinase activity and may be useful in the treatment of diabetes and other diseases or conditions characterized by impaired endogenous insulin production or an impaired response to endogenous insulin.

Abstract: Methods for the detection of creatine compound levels in body fluid samples are discussed. Portable kits capable of determining creatine levels using non-invasive and visually detectable methods are also included.

Abstract: The invention provides isolated nucleic acids molecules, designated Kinase and Phosphatase nucleic acid molecules, which encode novel protein kinase and protein Phosphatase polypeptides. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing Kinase and Phosphatase nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a Kinase and Phosphatase gene has been introduced or disrupted. The invention still further provides isolated Kinase and Phosphatase proteins, fusion proteins, antigenic peptides and anti-Kinase and Phosphatase antibodies. Diagnostic, screening, and therapeutic methods utilizing compositions of the invention are also provided.

Abstract: A stabilized reagent for the photometric enzymatic determination of creatine kinase (CK) in biological sample material by forming ATP from creatine phosphate and ADP and detecting the ATP formed, containing an organic or inorganic sulphur compound preferably in a submolar amount relative to the added CK activator. A corresponding reagent in a liquid form is stable at 2 to 8° C. for up to 12 months without significant loss of function.

Abstract: Described herein are methods which identify candidate agents as binding to a protein or as a modulator of the binding characteristics or biological activity of a protein. Generally, the methods involve the use of ADP or phosphate. The assays can be used in a high throughput system to obviate the cumbersome steps of using gels or radioactive materials.

Abstract: Disclosed is a stabilized enzyme composition for use in clinical examination, comprising: (a) an enzyme component comprising at least two enzymes selected from the group consisting of alkaline phosphatase, creatine kinase and alanine aminotransferase; (b) a stabilizer component comprising effective stabilizing amounts of an albumin, and at least one saccharide selected from the group consisting of trehalose and sorbitol; and (c) an aqueous medium having dissolved therein the components (a) and (b). The enzyme composition of the present invention is stable for a prolonged period of time not only under non-freeze refrigeration conditions, but also under freezing conditions or under conditions for non-freeze refrigeration after thawing of the frozen composition, as compared to the conventional enzymatic compositions.

Abstract: Amounts of components in a specimen can be analyzed with excellent quantitativity. The analysis includes: measuring an amount of a component to be analyzed in a specimen; measuring an amount of a standard component present originally and homeostatically in the specimen other than the component to be analyzed; determining the amount of the specimen from the amount of the standard component thus measured and a known concentration of the standard component in the specimen; and determining a concentration of the component to be analyzed in the specimen from the amount of the specimen thus determined and the amount of the component to be analyzed thus measured. The quantitative analysis of the present invention allows a component to be analyzed to be measured with high quantitativity as shown in FIG. 1.

Abstract: A biosensing system and method are disclosed for the quantitative determination of the concentration of particular analyte ions in biological sera in the presence of interfering ion species and, more particularly, to the quantitative determination of the concentration of analytes that are produced by biologically active materials including enzyme catalyzed reactions and are indicative of the presence of reactant species of interest in blood. The invention further deals with interfering ion species in a manner that eliminates the need for additional sensors or separate baseline sensors. The invention is exemplified by embodiments for the determination of the concentration of blood urea and creatinine using an ion transport related time delay potentiometric determination technique.

Abstract: A liquid reagent for detecting creatine kinase activity. The liquid reagent includes G6PDH, hexokinase, creatine phosphate, glucose, ADP, NAD(P) and thioglycerol, which are split into two separate batches of reagents for storage. G6PDH is contained in one batch, and thioglycerol together with a further substance is contained in the other batch. The batches of reagents are mixed together prior to use for detection purposes.

Abstract: In accordance with the present invention, fibroproliferative disease or condition characterized by such symptoms as increased levels of c-Jun homodimers, increased heterodimerization of c-Jun with another signaling peptide, increased levels of phosphorylated c-Jun, or increased presence of Jun kinase are treated by administering to the subject an amount of a compound effective to ameliorate one or more of the symptoms of the disease or condition, for example, an antiproliferative or antifibrotic agent. Preferred compounds for administration according to the invention are antisense c-Jun oligonucleotides and compounds that block c-Jun phosphorylation, such as pentoxifylline, or a functional derivative or metabolite thereof. Also provided by the present invention are in vitro tests for identifying whether a test compound is useful for treatment of a subject afflicted with such a disease and kits useful for conducting such assays.

Abstract: This invention discloses methods for detecting specific nucleic acid sequences, interrogating the identity of a specific base within a sequence, and assaying endonuclease and exonuclease activity. DNA or RNA probes are hybridized to target nucleic acid sequences. Probes that are complementary to the target sequence at each base are depolymerized, while probes which differ from the target at the interrogation position are not depolymerized. The nucleic acid detection systems utilize the pyrophosphorolysis reaction catalyzed by various polymerases to produce deoxyribonucleoside triphosphates or ribonucleoside triphosphates. dNTPs are transformed to ATP by the action of NDPK. The ATP produced by these reactions is detected by luciferase or NADH based detection systems.

Abstract: Test strips by which creatine kinase activity can be quantitatively measured at a high sensitivity within a broad measuring range and which have excellent storage stability. Particularly, test strips for the measurement of creatine kinase activity, which comprises a carrier, a dehydrogenase, a diaphorase, NAD or NADP, and a water-soluble tetrazolium compound.

Abstract: The present invention relates to an enzyme sensor for measuring the concentration or activity of an analyte in a test fluid. The sensor has at least one enzyme layer comprising an immobilized enzyme for which the analyte is a substrate. The immobilized enzyme is obtained by formation of one or more covalent link(s), optionally by using a cross-linking agent, between the enzyme and at least one type of macromolecule in the presence of a competitive inhibitor for said enzyme. The present invention also relates to a membrane for an enzyme sensor. Furthermore, the invention relates to a method for stabilizing the enzymatic activity of an enzyme sensor.

Abstract: The present invention relates to screening methods for the identification of compounds and compositions useful as novel antibiotics and antibacterial agents. In particular, the present invention relates to methods utilizing two-component regulatory switches which includes regulatory switches comprising a prokaryotic enzyme. Histidine protein kinase is a regulatory switch that is activated to autophosphorylate by a signal transduction mechanism. The invention also relates to methods of identifying inhibitors of enzyme activity particularly in bacterial cells. A high-throughput assay system useful in the large-scale screening of protein kinase inhibitors is also provided. The invention further provides the phosphorylation of SpoOF including a histidine.

Abstract: A dry analytical element for quantitatively analyzing creatine kinase MB isozyme is composed of a support and a creatine kinase MB isozyme-detecting agent layer. The detecting layer contains an antibody which specifically inhibits activity of M sub-unit of creatine kinase, creatine phosphate and adenosine diphosphate which react with each other to give adenosine triphosphate in the presence of creatine kinase MB isozyme, a buffer compound which keeps the layer in the range of pH 5.5 to pH 8.5 for the formation of adenosine triphosphate, and an indicator composition which reacts with the formed adenosine triphosphate to give a spectroscopically detectable compound, said indicator composition having hexokinase, nicotinamide-adenine dinucleotide or its phosphate in an oxidized form, and glucose-6-phosphate dehydrogenase but not containing glucose in such amount as to give the spectroscopically detectable compound.

Abstract: In accordance with the present invention, it has been discovered that monocyte conditioned medium (MCM) obtained from patients with liver disease stimulates the proliferation of fibroblasts. Platelet derived growth factor (PDGF) has also been found to stimulate fibroproliferation of fibroblasts, and to be at least partially responsible for the fibroproliferative effect of the MCM. Further in accordance with the present invention, the effect of MCM and PDGF on the expression of c-fos and c-jun has been investigated, because c-fos and c-jun form AP-1 complexes which can stimulate genes involved in proliferation. It has recently been reported that pentoxifylline inhibits platelet derived growth factor-stimulated proliferation. The mechanism of this action of pentoxifylline is unclear. Thus, in the course of the work undertaken as part of the present invention, studies were conducted to determine whether pentoxifylline altered the expression of c-fos and c-jun.

Abstract: A dry analytical element for quantitatively analyzing creatine kinase is composed of a support and a creatine kinase-detecting agent layer. The creatine kinase-detecting agent layer contains creatine phosphate and adenosine diphosphate which react with each other to give adenosine triphosphate in the presence of creatine kinase, a buffer compound having a sulfonic acid group which keeps the layer in the range of pH 5.5 to pH 8.5 for the formation of adenosine triphosphate, and an indicator composition which reacts with the formed adenosine triphosphate to give a spectroscopically detectable compound. An analytical element for analysis of creatine kinase MB isozyme further contains in the layer an antibody which specifically inhibits activity of M sub-unit of creatine kinase.

Abstract: Specific binding ligands can be detected with an assay which utilizes an immobilized receptor for the ligand, an immobilized reporter enzyme, an inhibitor antibody and a a water-soluble conjugate of the ligand and an anti-inhibitor antibody. Both antibodies are specific for the reporter enzyme, but the antibodies affect enzymatic activity differently. The inhibitor antibody effectively shuts down the activity of the reporter enzyme when it is complexed thereto. The anti-inhibitor antibody binds to the reporter enzyme, does not affect the enzymatic activity, but prevents the binding of the inhibitor enzyme. This assay provides a direct correlation of the generated signal to the target specific binding ligand.

Abstract: A method for determining adenosine 5' diphosphate (ADP) contained in a liquid sample by means of an enzymatic reaction, comprising reacting the sample at 15 to 45.degree. C. at least in the presence of glucose, ADP-dependent hexokinase, and oxidized NAD(P), a glucose-6-phosphate dehydrogenase, and one or more salts releasing ions selected among magnesium, cobalt, and manganese ions and then determining the ADP contained in the sample together with the AMP resulting from the reaction based on the amount of the reduced NAD(P) yielded. This method has advantages in that the limit of determination is high because ADP is determined based on the amount of the reduced NAD(P) yielded, and since the reduced NAD(P) has a definite molecular extinction coefficient, the value found is highly reliable and uninfluenced by the reducing substances contained in the sample.

Abstract: An immunoassay is provided which is selective for an analyte over immunologically related substances which may be present in a sample to be tested. The presence of the analyte is detected using a first binding substance, typically an antibody, in the presence of a second binding substance, typically another antibody. The first binding substance recognizes an epitope which is characteristic of the analyte and cross-reacts with a related epitope on the cross-binding substance. The second binding substance preferentially binds the common epitope on the cross-binding substance, thus reducing non-specific binding of the first binding substance.

Abstract: A method for avoiding influence of hemoglobin, in which the influence is caused by the changes with time in the absorption wavelength of hemoglobin when light absorption of a color reaction of a sample is measured by a rate assay, wherein the method comprises measuring the light absorption at a wavelength of from 517 to 529 nm or from 580 to 592 nm.

Abstract: A TRAF2 (Tumor Necrosis Factor receptor Associated Factor-2) kinase and DNA encoding it are described. The invention provides assays employing the TRAF2 kinase which are useful to identify candidate modulators of TRAF2-dependent signaling pathways.

Abstract: The invention relates to a method of identifying a compound capable of disrupting the addition of choline onto a bacterial cell surface component comprising incubating a sample of bacteria in a solution containing choline in the presence or absence of a test compound, and assessing the effect of the test compound on the addition of choline onto the bacterial cell surface component, wherein a lower level of choline on the cell surface component in the presence of the test compound, compared with the level of choline on the cell surface component in the absence of the test compound, is an indication that the test compound inhibits the addition of choline onto the cell surface component.

Abstract: The present invention relates to methods useful for identifying compounds capable of specifically regulating actin polymerization, stress fiber formation or focal adhesion assembly by regulating G.sub..alpha.12 and/or G.sub..alpha.13 activity in cells involved in inflammatory responses, immune responses, allergic responses and neuronal responses, kits to perform such assays and methods to control disease related to such responses.

Abstract: A reagent for assaying creatine kinase, which contains glucokinase or hexokinase, glucose-6-phosphate dehydrogenase, adenosine 5'-diphosphate, creatine phosphate, oxidized nicotinamide adenine dinucleotide phosphate, magnesium salt, and glucose; and which comprises a first liquid reagent having a pH value of 7.5 to 10 and containing at least glucokinase and/or hexokinase, glucose-6-phosphate dehydrogenase, adenosine 5'-diphosphate and creatine phosphate, and a second liquid reagent having a pH value of 2 to 5 and containing at least oxidized nicotinamide adenine dinucleotide phosphate. The present reagent can be stably stored in the form of liquid for a long period of time at normal or low temperature in dark or light place. Therefore, it can be used without dissolution procedure after being preserved for a long term at the site of clinical examination and is also applicable to automatic analyzers.

Abstract: The object of the invention is to provide a reagent for quantitatively determining creatine kinase with improved storability in liquid form as well as a method for quantitatively determining creatine kinase with stable measurements. Disclosed are a method for quantitatively determining creatine kinase, which comprises activating creatine kinase in a sample in an aqueous medium in coexistence with a trivalent phosphorus compound and a sulfhydryl-containing compound and then determining creatine kinase activity; a method for stabilizing a sulfhydryl-containing compound, which comprises allowing a trivalent phosphorus compound to coexist with a sulfhydryl-containing compound; and a reagent for quantitatively determining creatine kinase, which comprises a trivalent phosphorus compound, a sulfhydryl-containing compound, and a reaction substrate for creatine kinase.

Abstract: The present invention provides such a measuring method of the CK isoform and a reagent therefor. Accordingly, the present invention provides a reagent for measuring creatine kinase (CK) activity comprising an antibody, wherein said antibody inhibits CK-M.sub.T subunit, but does not inhibit CK-M.sub.S subunit.The present invention also provides a method for measuring CK activity comprising determining an inhibition of CK of body fluids using a reagent for measuring CK activity, wherein said reagent comprises an antibody which inhibits CK-M.sub.T subunit, but does not inhibit CK-M.sub.S subunit.

Abstract: Kits and methods for measuring enzyme activities and metabolites using NADH and NADPH analogs are disclosed. The analogs have extended stability in aqueous solutions and can used as replacements for NADH or NADPH cofactors in analytical procedures. Preferred aspects of the invention include kits containing the NADH and NADPH analogs for use in the measurement of ALT activity, AST activity, Urea, Ammonia, Salicylate, Triglycerides, Pyruvic Acid, Sorbitol Dehydrogenase activity, 5'-Nucleotidase activity, Creatine Kinase activity, 2,3-Diphosphoglyceric Acid, Adenosine 5'-triphosphate, .alpha.-Hydroxybutyrate Dehydrogenase activity, Lactate Dehydrogenase activity and the Carbon Dioxide content in analytical samples.

Abstract: Disclosed is a method for stably storing a liquid diagnostic reagent, comprising air-hermetically keeping the liquid diagnostic reagent in a closed container in the presence of a disoxidant therein. Preferably, at least one of the liquid diagnostic reagent and the disoxidant is covered with a separating container made of a material pervious to oxygen but not to solutions. The liquid diagnostic reagent may comprise an enzyme or an indicator.

Abstract: Materials can be assayed for activity as an inhibitor of post-translational protein phosphorylation by adding the material to a growing culture of a prokaryotic organism such as a streptomycete; allowing the culture to grow for a period of time in the presence of the material; and observing the culture for altered development relative to development of the prokaryotic organism grown in the absence of the material. Observation of altered development is indicative that the material has activity as an inhibitor of post-translational protein phosphorylation. In particular, the material to be tested can be added to a growing culture of the prokaryotic organism by placing a carrier disk bearing the material on a freshly seeded plate. Inhibition of the development of aerial mycelia and spore formation is an indicator that the material has activity as an inhibitor of post-translational protein phosphorylation.

Abstract: The present invention relates to screening methods for the identification of compounds and compositions useful as novel antibiotics and antibacterial agents. In particular, the present invention relates to methods utilizing two-component regulatory switches, for example, those comprising a prokaryotic enzyme such as histidine protein kinase that is activated to autophosphorylate by a signal transduction mechanism. The invention also relates to methods of identifying inhibitors of enzyme activity, particularly in bacterial cells. In particular, a high-throughput assay system useful in the large-scale screening of protein kinase inhibitors is disclosed.

Abstract: A stable two-part liquid reagent composition for measuring creatine kinase activity contains, in the first part, adenosine 5'-adiphosphate, kexokinase or glucokinse, nicotinamide adenine dinucleotide (phosphate), glucose-6-phosphate dehydrogenase, and thioglycerol, 2-mercaptoethanol, or 2-mercaptoethanesulfonic acid or a salt thereof; and contains, the second part, creatine phosphate at a pH of 7.5 to 10.0. Glucose and magnesium ions can be included in either part or in both parts. The two-part composition is stable even after being stored for 3 months at 10.degree. C.