Incubation of cultured bovine adrenal medullary cells with p-chloromercuribenzoate (50-500 microM), a sulfhydryl-reacting agent, caused an increase in the secretion of catecholamines, p-Chloromercuriphenyl sulfonate, a p-chloromercuribenzoate analogue that poorly penetrates the cell membrane, caused a similar increase in catecholamine secretion. In both cases, catecholamine secretion was dependent on extracellular Ca2+. Furthermore, p-chloromercuribenzoate caused both 45Ca2+ influx into the cells and an increase in the intracellular free Ca2+ concentration. The increases in catecholamine secretion and 45Ca2+ influx behaved similarly in relation to p-chloromercuribenzoate concentration. The time courses of the increased secretion, 45Ca2+ influx, and intracellular free Ca2+ concentration by p-chloromercuribenzoate were also quite similar. The stimulation of catecholamine secretion by p-chloromercuribenzoate was reversed by washing the cells with dithiothreitol-containing medium, but not by dithiothreitol
1GGV: Structure of the C123S mutant of dienelactone hydrolase (DLH) bound with the PMS moiety of the protease inhibitor phenylmethylsulfonyl fluoride (PMSF).
Yves, You dont say whether or not you are briefly fixing the sections. The binding of the b-galactosidase to lysosomes is relatively loose, and that may contribute to what you are seeing. Brief fixation in paraformaldehyde vapor may suffice. Also, something worth considering, have you tried inhibiting the enzyme with e.g. D-galactonolactone, lactose and/or p-chloromercuribenzoate? This is only speculation as I have never come across this phenomenon personally, but, as with Ray, have never performed the technique on pancreas.=0D One way to get by the problem of diffusion artefact, if that is what this is, would be to employ a semi-permeable membrane enzyme =0Ahistochemical technique. Ronnie Houston, MS, HT(ASCP)QIHC Anatomic Pathology Manager Nationwide Childrens Hospital =0A700 Childrens Drive Columbus, OH 43205 (614) 722 5465 [email protected] Columbus Childrens Hospital is now Nationwide Childrens Hospital www.NationwideChildrens.org=0D -----Original Message----- ...
Anson, Fred C. and Shi, Chunnian and Steiger, Beat (1997) Novel Multinuclear Catalysts for the Electroreduction of Dioxygen Directly to Water. Accounts of Chemical Research, 30 (11). pp. 437-444. ISSN 0001-4842. https://resolver.caltech.edu/CaltechAUTHORS:20171122-083839178 Shi, Chunnian and Steiger, Beat and Yuasa, Makoto et al. (1997) Electroreduction of O_2 to H_2O at Unusually Positive Potentials Catalyzed by the Simplest of the Cobalt Porphyrins. Inorganic Chemistry, 36 (20). pp. 4294-4295. ISSN 0020-1669. https://resolver.caltech.edu/CaltechAUTHORS:20180523-110226758 Shi, Chunnian and Anson, Fred C. (1997) Electrochemical Assessment of the Stability of [Os(NH_3)_5(OH_2)]^(2+) in Aqueous Solutions. Inorganic Chemistry, 36 (12). pp. 2682-2687. ISSN 0020-1669. https://resolver.caltech.edu/CaltechAUTHORS:20180529-110854891 ...
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Substrate effects on the activation kinetics of Chinese hamster dihydrofolate reductase by p-chloromercuribenzoate (pCMB) have been studied. On the basis of the kinetic equation of substrate reaction in the presence of pCMB, all modification kinetic constants for the free enzyme and enzyme-substrate binary and ternary complexes have been determined. The results of the present study indicate that the modification of Chinese hamster dihydrofolate reductase by pCMB shows single-phase kinetics, and that changes in the enzyme activity and tertiary structure proceed simultaneously during the modification process. Both substrates, NADPH and 7,8-dihydrofolate, protect dihydrofolate reductase against modification by pCMB. In the presence of a saturating concentration of NADPH, the value of kcat for 7,8-dihydrofolate in the enzyme-catalysed reaction increased four-fold on modification of Cys-6, accompanied by a two-fold increase in Km for the modified enzyme. The utilization of the binding energy of a ...
1. The ring-fission of cis- and trans-acenaphthene-1,2-diol by rat liver microsomes was studied. 2. 1,8-Naphthalic acid was detected and isolated after microsomal incubations of the diols. 3. The accompanying reduction of NAD+ was followed spectrophotometrically. 4. The optimum pH for the microsomal reaction was 9·4 for the oxidation of the cis-diol and 9·8 for that of the trans-diol. 5. p-Chloromercuribenzoate and 2,4-dichlorophenol inhibited the reaction. 6. Possible mechanisms for the microsomal ring-fission, involving 1,8-naphthalic aldehyde, are discussed.. ...
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Cortiglioni, S.; Baralis, M.; Bernardi, G.; Boella, G.; Bonometto, S.; Bruscoli, M.; Carretti, E.; Cecchini, S.; Fabbri, R.; Gervasi, M.; Macculi, C.; Monari, J.; Musso, C.; Nicastro, L.; Orfei, A.; Peverini, O.; Poppi, S.; Sbarra, C.; Sironi, G.; Tascone, R.; Tucci, M.; Vinyajkin, E.; Zannoni, M ...
TY - JOUR. T1 - Cobalt porphyrin catalyzed reduction of CO2. Radiation chemical, photochemical, and electrochemical studies. AU - Behar, D.. AU - Dhanasekaran, T.. AU - Neta, P.. AU - Hosten, C. M.. AU - Ejeh, D.. AU - Hambright, P.. AU - Fujita, Etsuko. PY - 1998/4/23. Y1 - 1998/4/23. N2 - Several cobalt porphyrins (CoP) have been reduced by radiation chemical, photochemical, and electrochemical methods, in aqueous and organic solvents. In aqueous solutions, the CoIP state is stable at high pH but is shorter lived in neutral and acidic solutions. Stable CoIP is also observed in organic solvents and is unreactive toward CO2. One-electron reduction of CoIP leads to formation of a species that is observed as a transient intermediate by pulse radiolysis in aqueous solutions and as a stable product following reduction by Na in tetrahydrofuran solutions. The spectrum of this species is not the characteristic spectrum of a metalloporphyrin π-radical anion and is ascribed to Co0P. This species binds ...
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View Notes - General Properties of Viruses-review from MCB 2000 at University of Florida. General Properties of Viruses Structure 1. Nucleic acid - Single or double stranded. Segmented or
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An intracellular β-glucosidase (EC 3.2.1.21) from Debaryomyces hansenii Y-44 was purified and characterized. The enzyme was purified to homogeneity from the cell-free extracts by a combination of column chromatography with DEAE-Toyopearl 650M and Butyl-Toyopearl 650S. The size of the enzyme was 95 kDa by native-PAGE, and the pl was 4.9. The enzyme was the most active at pH 7.0 and at around 25°C. The activity was highly tolerant to glucose, and only 20% inhibited in 500 mM glucose. The enzyme was tolerant to ethanol; its activity was only slightly reduced by 6% in the presence of 15% (v/v) ethanol. As the enzyme was inhibited with p-chloromercuribenzoate, it belongs to the class of SH-enzymes. The enzyme efficiently released monoterpenols from the glycosides extracted from Muscat grape must. The fermentation of Muscat juice coupled with the enzyme addition produced a considerable increase in the concentration of monoterpenols. Especially the linalool and nerol contents increased by 90% and ...
Atlassian Sourcetree is a free Git and Mercurial client for Windows. Atlassian Sourcetree is a free Git and Mercurial client for Mac. ...