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61.
Surface plasmon resonance (SPR) has been successfully applied for the simple, rapid, and label-free assay of various biomolecules. This assay evaluates a novel wavelength modulation SPR biosensor for the detection of tetanus toxin. The wavelength modulation SPR biosensor is designed based on fixing the incident angle of light and measuring the reflected intensities in the resonance wavelength range spanning 400-800 nm simultaneously. Tetanus toxin (TeNT), one of the most potent toxins known, is synthesized as a 150 kDa single polypeptide chain. The SPR biosensor has been shown to be capable of directly detecting concentration of tetanus toxin as low as 0.028 Lf ml−1. Under selected experimental conditions, the SPR biosensor has a good reproducibility, sensitivity and reversibility. The results illustrate how wavelength modulation SPR biosensor can be used to detect biomolecular interactions. 相似文献
62.
J. Sereikaité D. Iljasevičiené G. Dienys H. Danilčenko V. Gavrilova 《Applied biochemistry and biotechnology》1993,43(2):153-160
Ascorbate oxidase fromCucurbita sp. was isolated by ammonium sulfate precipitation and DEAE-dextran-silochrome column chromatography. The thermal and pH stabilities
of the purified enzyme were investigated. TheK
M forl-ascorbic acid (1.5 mM) and chlorohydroquinone (0.37 mM) was determined. Substrate specificity of ascorbate oxidase was investigated and compared with those of laccases fromCoriolus hirsutus andCerrena maxima. Ascorbate oxidase was covalently bound to a polymeric membrane and used in an enzyme electrode for ascorbic acid. 相似文献
63.
Darko P. Aanin Sneana Rajkovi Dora Molnar-Gabor Milos I. Djuran 《Monatshefte für Chemie / Chemical Monthly》2004,135(12):1445-1453
Summary. Hydrolytic reactions between various palladium(II) complexes of the type cis-[Pd(L)(H2O)2]2+ in which L is ethylenediamine (en), 1,2-propylenediamine (1,2-pn), isobutylenediamine (ibn), 1,2-diaminocyclohexane (1,2-dach), N-methylethylenediamine (Meen), N,N,N,N-tetramethylethylenediamine (Me4en), S-methyl L-cysteine (MeS-L-HCys), L-methionine (L-HMet), and 2,5-dithiahexane (dth) and dipeptide N-acetylated L-methionylglycine (MeCOMet-Gly) were studied by 1H NMR spectroscopy. The reactions were carried out in the pH range 2.0–2.5 and at 50°C. In all these reactions, palladium(II) complex bound to a methionine residue effects the regioselective cleavage of the amide bond involving the carboxylic group of methionine. We found that the rate of hydrolysis and mechanism of this reaction are strongly dependent from the nature of the chelate ligand L in palladium(II) complexes of the type cis-[Pd(L)(H2O)2]2+. 相似文献
64.
Cartoni C Schininà ME Maras B Nonno R Vaccari G Di Baria MA Conte M Liu QG Lu M Cardone F Windl O Pocchiari M Agrimi U 《Journal of chromatography. A》2005,1081(1):122-126
Cerebral formation of the pathological isoform of the prion protein (PrP) is a crucial molecular event in prion diseases. The bank vole (Clethrionomys glareolus) is a rodent species highly susceptible to natural scrapie. The PrP gene of bank vole is polymorphic (Met/Ile) at codon 109. Here we show that homozygous 109Met/Met voles have incubation times shorter than heterozygous 109Met/Ile voles after experimental challenge with three different scrapie isolates. An HPLC-MS/MS method was optimized and applied to investigate whether in heterozygous animals both PrP allotypes are able to undergo pathological conversion. The results demonstrate that both allotypes of the prion protein participate to pathological deposition. 相似文献
65.
A rapid microtiter plate assay for the detection of inhibitors of the Na+, K+-ATPase has been developed. The assay is based on the measurement of inorganic phosphate released from the substrate, ATP,
and has been designed to be carried out in the individual wells of a microtiter plate. Since the production of inorganic phosphate
is determined colorimetrically, multiple samples can be tested simultaneously using a microtiter plate reader. This microtiter
plate assay is particularly useful for screening large numbers of samples, such as microbial culture supernatants. 相似文献
66.
Toshiyuki Chikuma Yoko ShimabukuroTakeya Iguchi Akira TanakaKyoji Taguchi Takeshi KatoMitsune Yamaguchi Hiroshi Hojo 《Analytica chimica acta》2002,457(2):157-164
A rapid and sensitive assay for the determination of Dns-His-Lys-Arg-His-Lys cleaving enzyme activity is reported. This assay is based on fluorimetric detection of a dansylated dipeptide, 5-dimethylaminonaphthalene-1-sulfonyl-His-Lys, enzymatically formed from the substrate 5-dimethylaminonaphthalene-1-sulfonyl-His-Lys-Arg-His-d-Lys, after separation by high-performance liquid chromatography (HPLC) using a C-18 reversed-phase column by isocratic elution. This assay is sensitive enough to measure 5-dimethylaminonaphthalene-1-sulfonyl-His-Lys at concentrations as low as 7 pmol, and yields highly reproducible results and requires less than 9.0 min per sample for separation and quantitation. The optimum pH for Dns-His-Lys-Arg-His-Lys cleaving enzyme activity was 7.5-8.0. The Michaelis constant (Km) and the maximum velocity (Vmax) values were 33.3 μM and 47.07 pmol/(μg h), respectively with the use of enzyme extract obtained from bovine pituitary. By using this assay, axonal transport of this enzyme activity was observed 48 h after double ligations of rat sciatic nerves. The high sensitivity and selectivity of this assay would be useful for clarification of the physiological role of this enzyme. 相似文献
67.
Transition state analogues (TSAs) have long been regarded as ideal templates for the preparation of catalytically active synthetic imprinted polymers. In the current work, however, a new type of molecularly imprinted polymer (MIP) was synthesized with the substrate (homovanillic acid, HVA) as the template and hemin introduced as the catalytic center, with the use of plural functional monomers to prepare the active sites. The MIP successfully mimicked natural peroxidase, suggesting that it may not be imperative to employ a TSA as the template when preparing enzyme-like imprinted polymers and that the imprinted polymer matrix provided an advantageous microenvironment around the catalytic center (hemin), essentially similar to that supplied by apo-proteins in natural enzymes. Significantly, by taking advantage of the special structure of hemin and multiple-site interactions provided by several functional monomers, the intrinsic difficulties for MIPs in recognizing template molecules in polar solutions were overcome. The newly developed polymer showed considerable recognizing ability toward HVA, catalytic activity, substrate specificity and also stability, which are the merits lacked by the natural peroxidase. Meanwhile, the ease of recovery and reuse the MIP implies the potential for industrial application. 相似文献
68.
InteractionbetweenMetalinMetalloEnzymeandSmallBiologicalMolecules¥HuJie-Han;ShuZan-Yong;TaoLi-Mei;ChengGuo-Bao(DalianInstitut... 相似文献
69.
Mai Thanh Thi Nguyen 《Tetrahedron letters》2004,45(46):8519-8522
A novel dimeric methanodibenzoxocinone, named neosappanone A (1), possessing a unique unprecedented novel carbon framework, has been isolated from the heartwood of Caesalpinia sappan L. of Vietnam, and its structure was elucidated on the basis of spectroscopic analysis. Neosappanone A (1) competitively inhibited xanthine oxidase in a concentration-dependent manner (IC50, 29.7 μM; Ki, 16.3 μM). 相似文献
70.