Chromatography of beta-glucuronidase from bovine liver. A study of the enzyme binding sites of prepared adsorbents.

beta-Glucuronidase from bovine liver was adsorbed to the adsorbents prepared with CH-Sepharose 4B and either the competitive inhibitor or its analogs such as p-aminophenyl 1-thio-beta-D-glucuronic acid, -glucoside, -galactoside, and N-acetyl glucosaminide. The adsorbed enzyme was eluted at 0.1 or 0.5 M NaCl by a stepwise gradient. Chromatography of the enzyme was also performed by using the adsorbents prepared with Epoxy-activated Sepharose 6B and amine compounds or other compounds. In order to see whether the hydroxyl groups of the sugar parts in the ligand are necessary for the adsorption of the enzyme, chromatography was performed by using the adsorbents prepared with sugar derivatives as the ligand. As a result, it was found that beta-glucuronidase had an affinity for adsorbents prepared with either acetyl derivatives or methoxy derivatives of glycosides and CH-Sepharose 4B. From the results of elution of the enzyme with NaCl from adsorbents having amide bonding, it was clarified that the affinity of the enzyme for adsorbents without glycosides in the ligands correlated with acidity of the amide in the adsorbents. Hydrogen bond chromatography was performed with the prepared adsorbents. The enzyme was adsorbed under a high concentration of ammonium sulfate, and the elution of the adsorbed enzyme from adsorbents was examined by the degradation of salt. The enzyme was most easily eluted from aminoethyl 1-thio-beta-D-glucuronic acid-CH Sepharose 4B at 0.9 M ammonium sulfate and at 0.5 M concentration of the salt with p-aminophenyl 1-thio-beta-D-glucuronic acid-CH Sepharose 4B.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interaction between dyes and polyelectrolytes. X. Excitation energy transfer in cationic dye–polyanion system

The quantum yield for the trypaflavine-photosensitized oxidation of 1-(methylthio)ethyl-3-carbamoylpyridinium chloride to 1-(methylsulfinyl)ethyl-3-carbamoylpyridinium chloride increased with increase in the concentration of methylene blue added. It was inferred that the increase in the quantum yield is due to the excitation energy transfer from trypaflavine to methylene blue. The efficiency of excitation energy transfer was enhanced on addition of potassium poly(vinyl sulfate) and was dependent on the polyanion/dye ratio. The efficiency of excitation energy transfer at the most appropriate polyanion/dye ratio was about 5 times as high than that in the absence of polyanion. The binding of dye to potassium poly(vinyl sulfate) was investigated spectrophotometrically. Correlation with the dye binding to potassium poly(vinyl sulfate) and the efficiency of excitation energy transfer between dyes was discussed.     

Modification of intracellular Ca2+ dynamics by laser inactivation of inositol 1,4,5-trisphosphate receptor using membrane-permeant probes

A membrane-permeant malachite green-conjugated IP3 analog (MGIP3/PM) was synthesized as a probe for small molecule-based CALI (smCALI), and its effect on the Ca2+ signaling in intact DT40 chicken B cells was examined. In DT40 B cells treated with the smCALI probe, laser irradiation inhibited IP3-induced Ca2+ oscillations in response to B cell receptor stimulation, demonstrating that IP3R was acutely inactivated. We then applied smCALI to clarify the mechanism of capacitative Ca2+ entry (CCE), in which involvement of IP3R has been suggested. Despite the inactivation of IP3R by smCALI, thapsigargin-induced CCE remained unaffected, providing evidence that functional IP3R is not required for CCE in DT40 cells. These results demonstrate the potency of the smCALI technique for the study of the roles of IP3R in complex intracellular Ca2+ dynamics.     

Preparation of ternary platinum(II) complexes with N-(omega-phenylalkyl)-1,2-ethanediamine and 2,2'-dipyridine and the effect of the methylene chain length of the N-(omega-phenylalkyl)-1,2-ethanediamine in the complexes on intermolecular interactions with various arylsulfonates

A series of ternary complexes comprised of platinum(II), 2,2'-dipyridine, and N-(omega-phenylalkyl)-1,2-ethanediamine was prepared by varying the number (n) of methylene chain carbons between the phenyl group and one of the amino groups of 1,2-ethanediamine. NMR measurements indicated that intramolecular stacking occurred for n=1 and intermolecular stacking occurred for n=3 for several of the aryl sulfonates.     

Structure of gentiodelphin,an acylated anthocyanin isolated from Gentiana Makinoi, that is stable in dilute aqueous solution

Structure of gentiodelphin is determined to be 5, 3′-d8-O-(6-O-$\text{trans}$-caffeoylβ-D-glucosyl)-3-O-(β-D-glucosyl)delphinidin. The anthocyanin is stable in dilute neutral aqueous solution. This stabilization may be caused from intramoleculaur hydrophobic interactions among the aromatic nuclei; the anthocyanidin being sandwiched win between two caffeic acids.     

Photo-thermal properties of poly(4-tert-butoxycarbonyloxystyrene) bearing imino sulfonate units

Copolymers of 4-tert-butoxycarbonyloxystyrene (BOS) and 9-fluorenilideneimino p-styrenesulfonate (FISS) were synthesized. FISS units in copolymers became p-styrenesulfonic acid units upon ultraviolet irradiation. The irradiated copolymers thermally decomposed to poly(hydroxystyrene) by liberating tert-butoxycarbonyl (BOC) units at temperatures where the unirradiated copolymers were stable. The thermal decomposition of the copolymers catalyzed by sulfonic acid formed photochemically was studied by thermogravimetry. The pseudo first-order rate constant (k) and the activation energy for the acid catalyzed thermal decomposition of BOC units in copolymers were evaluated. The thermolysis of the irradiated copolymer system was compared with that of the irradiated blended system of poly(4-tert-butoxycarbonyloxystyrene) (PBOCST) and 9-fluorenilideneimino p-toluene-sulfonate (FITS). © 1993 John Wiley & Sons, Inc.