- Versatile small molecule kinase assay through real-time, ratiometric fluorescence changes based on a pyrene-DPA-Zn2+complex
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A real-time kinase assay method based on a ratiometric fluorescence probe that can be applied to various small-molecule kinases is described herein. The probe can trace the reversible interchange of ATP and ADP, which is a common phenomenon in most small-molecule kinase reactions, by a ratiometric fluorescence change. This property facilitates the monitoring of phosphorylation and dephosphorylation in small-molecule kinases, whereas most of the existing methods focus on one of these reactions. To prove the applicability of this method for small-molecule kinase assays, hexokinase and creatine kinase, which phosphorylate and dephosphorylate substrates, respectively, were analyzed. The ratiometric fluorescence change was correlated with the enzyme activity, and the inhibition efficiencies of the well-known inhibitors,N-benzoyl-d-glucosamine and iodoacetamide, were also monitored. Notably, the change in fluorescence can be observed with a simple light source by the naked eye.
- Kim, Jihoon,Oh, Jinyoung,Han, Min Su
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p. 10375 - 10380
(2021/03/23)
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- Role of metal cations and oxyanions in the regulation of protein arginine phosphatase activity of YwlE from Bacillus subtilis
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Protein arginine phosphorylation (pArg) is a relatively novel posttranslational modification. Protein arginine phosphatase YwlE negatively regulates arginine phosphorylation and consequently induces the expression of stress-response genes that are crucial for bacterial stress tolerance and pathogenic homolog Staphylococcus aureus virulence. However, little is known about the factors that affect the enzymatic activity of YwlE with the exception of the effect of oxidative stress. Herein, based on the hydrolysis of the chromogenic substrate p-nitrophenyl phosphate (pNPP) by YwlE, we investigate the role of metal cations and oxyanions in the regulation of YwlE activity. Interestingly, among the various cations that we tested, Ca2+ activates YwlE, while other cations, including Ag+, Co2+, Cd2+, and Zn2+, are inhibitory. Furthermore, as chemical analogues of phosphate, oxyanions play multiple roles in phosphatase activity. The regulatory switch Cys within the catalytic site regulates YwlE activity. Specifically, the thiol of this Cys could be alkylated by IAM (iodoacetamide) or oxidized by H2O2, resulting in enzymatic inhibition. Conversely, reducing reagents, such as DTT (dithiothreitol), β-me (β-mercaptoethanol), and TCEP (tris(2-carboxyethyl)phosphine) enhance YwlE activity. Additionally, as a stable analogue to pArg, pAIE binds to YwlE with a Kd of 149.1 nM and a binding stoichiometry n of 1.2 and inhibits YwlE with an IC50 of 316.3 ± 12.73 μM. The inhibition and activation of YwlE may have broad implications for the physiology, pharmacology and toxicology of metal cations and oxyanions.
- Huang, Biling,Huang, Chenyang,Huang, Shaohua,Liao, Xinli,Liu, Yan,Zhang, Yumeng,Zhao, Mingxiao,Zhao, Yufen,Zhao, Zhixing
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- Synthetic method of creatine
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The invention discloses a synthetic method of creatine. The method includes the steps of: adding water into a stainless steel reaction kettle having stirring, heating and reflux devices, adding guanidinoacetic acid and a catalyst AlCl3-K2CO3, adding dimethyl carbonate with stirring, heating the reaction kettle to increase the temperature therein, continuously performing the reaction, cooling the reaction product after the reaction is finished to separate creatine crystal out, and filtering and drying the creatine crystal to prepare the creatine. The synthetic method is less in reaction steps, is simple in process operation, is safe and environment-friendly and is greatly improved in product purity.
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Paragraph 0020
(2016/10/08)
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- Stable aqueous compositions comprising amide-protected bioactive creatine species and uses thereof
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The present invention provides amide-protected creatine molecules and compositions, containing one or more bioactive forms of creatine in aqueous compositions, wherein bioactive forms of creatine do not appreciably degrade into creatinine. Also provided are various beneficial effects of administering aqueous compositions having at least one amide-protected creatine molecule.
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- Process for Preparing Creatine, Creatine Monohydrate or Guanidinoacetic Acid
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A process for producing creatine, creatine monohydrate or guanidinoacetic acid is proposed, wherein firstly N-methylethanolamine or ethanolamine is catalytically dehydrogenated in each case in alkaline solution and the sarcosinate or glycinate solutions that are obtained in this manner are finally reacted under acidic conditions with a guanylating agent such as for example O-alkylisourea or cyanamide. In this manner products are obtained in high yields and very good purity where in contrast to the prior art no traces whatsoever of hydrocyanic acid, formaldehyde, chloroacetic acid or ammonia are present. The formation of the toxicologically critical dihydrotriazine is also avoided.
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Page/Page column 3
(2009/07/10)
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- Methods and compositions for diagnosing and treating arthritic disorders and regulating bone mass
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The present invention relates to improved diagnostic methods for early detection of a risk for developing an arthritic disorder in humans, and screening assays for therapeutic agents useful in the treatment of arthritic disorders, by comparing the levels of one or more indicators of altered mitochondrial function. Indicators of altered mitochondrial function include enzymes such as mitochondrial enzymes and ATP biosynthesis factors. Other indicators of altered mitochondrial function include mitochondrial mass, mitochondrial number and mitochondrial DNA content, cellular responses to elevated intracellular calcium and to apoptogens, and free radical production. Methods of treating, and of stratifying, human patients as such methods relate to disclosed indicators of altered mitchondrial function are also provided.
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Page/Page column 31
(2008/06/13)
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- PROCESS FOR THE SYNTHESIS OF GUANIDINE DERIVATIVES
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The invention pertains to a process for the synthesis of guanidine derivatives of Formula (I) wherein X is selected from OH, OM wherein M is an alkali or earth alkali metal ion, and NR1R2 wherein R1 and R2 are independently H, C1-C6 alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl, or C6-C10 aryl; which comprises a first step of reacting cyanogen chloride (N≡CCI) with NH3 in an organic solvent followed by a second step of reacting with an amine of Formula (II) wherein X has the previously given meaning, optionally followed by a step of converting a compound of Formula (I) into another compound of Formula (I).
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- Process for the preparation of creatine or creatine monohydrate
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A process for the preparation of creatine or creatine monohydrate by reaction of sodium or potassium sarcosinate with cyanamide at a temperature from 20 to 150° C. and a pH from 7.0 to 14.0 comprises carrying out the pH adjustment with carbonic acid.
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Page column 2-3
(2008/06/13)
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- Preparation of substituted guanidine derivatives
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Substituted guanidine derivatives of the formula I are prepared by reacting calcium cyanamide with a primary or secondary amino carboxylic acid or a primary or secondary amino sulfonic acid or their derivatives of the formula II where the substituents R1 and R2 have the meanings explained in the description.
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- Preparation of N-formamidinylamino acids from amino and formamidinesulfinic acids
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A practical synthetic procedure for the conversion of amino acids into N-formamidinylamino acids using formamidinesulfinic acid in basic water solution is presented.
- Jursic,Neumann,McPherson
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p. 1656 - 1658
(2007/10/03)
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- 6-[α-(ω-QUANIDINOALKANOYLAMIDO)ACYLAMIDO]PENICILLANIC ACIDS
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6-[α-(ω-GUANIDINOALKANOYLAMIDO)ACYLAMIDO]PENICILLANIC ACIDS, THE NON-TOXIC SALTS AND ESTERS THEREOF ARE USEFUL AS ANTIBACTERIAL AGENTS, THERAPEUTIC AGENTS IN ANIMALS, INCLUDING MAN, OF PARTICULAR VALUE AGAINST GRAM-NEGATIVE BACTERIA, AND AS ANIMAL FEED NUTRITIONAL SUPPLEMENTS.
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