Light-induced changes in resistivity of spinach chloroplasts toward modification with diazonium-1,2,4-triazole

Spinach chloroplasts in the light and in the dark were treated with several reagents for protein modification to see the effect of light on their resistivity toward modification. The reagents were p-diazobenzenesulfonic acid, diazonium-1-H-tetrazole, sodium-2,4,6-trinitrobenzenesulfonate, sodium-β-naphtoquinone-4,6-disulfonate and diazonium-1,2,4-triazole. No difference in the absorption spectrum was found between chloroplasts treated with these reagents in the light and those treated in the dark. However, these light- and dark-treated samples when solubilized with a nonionic detergent showed a difference in turbidity. Diazonium-1,2,4-triazole was the most suitable of the above reagents, and the solubilized sample of chloroplasts treated with diazonium-1,2,4-triazole in the light showed a turbidity which was about 2-fold higher than that of the same sample treated in the dark. This increase in turbidity was interpreted as being due to a change in the resistivity toward chemical modification of chloroplasts caused by illumination. In the presence of 3-(p-chlorophenyl)-1,1-dimethylurea, pentachlorophenol and 2-methylthio-4,6-bis-isopropylamino-s-triazine, which are inhibitors of the Hill reaction, the light-induced increase of turbidity was suppressed by 72, 78 and 62%, respectively. The addition of ATP caused a much greater increase of turbidity both in the light and in the dark. It was thus found that light and ATP induce a configurational change of chloroplasts or a conformational change of chloroplast proteins inside.     

Reaction of 3-amino-1:2:4-triazole with lactoperoxidase

Bovine lactoperoxidase (LPO) gradually lost its enzymatic activity when dialyzed against a solution of 3-amino-1:2:4-triazole (AT) and hydrogen peroxide at pH 7.0. Amino acid analysis of the completely inactive enzyme revealed the formation of a new ninhydrin-positive chromatographic peak. This peak which had been observed previously when catalases were similarly reacted with AT in the presence of hydrogen peroxide is attributed to the covalent reaction product of AT with a histidyl residue. Concurrently with the appearance of the new ninhydrin-positive peak, the histidine content of LPO decreased by approximately two residues. Four to five residues of tyrosine were also lost.     

Reaction of diazoalkanes with 1-substituted 2, 4-dioxopyrimidines. Formation of O2, N-3 and O4-alkyl products.

In non-aqueous solution, diazomethane and diazoethane react with the O2, O4 and N-3 sites of uridine, thymidine, 1-methyluracil and 1-methylthymine. Diazoethane has a higher affinity for alkylating oxygens than does diazomethane. The relative ratio of O2:O4:N-3 methyl products is 1:2:16 and of ethyl products the ratio is 1:1:2. When the diazoethane reaction is performed in neutral buffered solution, the same proportion of O2:O4:N-3 ethyl products is found, but the extent of reaction is very low. O2-alkylation greatly labilizes the glycosidic bond of thymidine and uridine toward acid hydrolysis. All O2 and O4 alkyl 1-substituted 2,4-dioxopyrimidines are dealkylated in weak acid but the O2 alkyl group is the more stable.     

Interaction of 2-n-heptyl-4-hydroxyquinoline-N-oxide with photosystem II in chloroplasts and subchloroplast particles

The effects of 2-n-heptyl-4-hydroxyquinoline-N-oxide on electron transport in thylakoids and oxygen-evolving photosystem II particles has been examined. Kinetic fluorescence studies reveal that the site of inhibition for alkyl derivatives of hydroxyquinoline-N-oxide (I50 approximately equal to 2 microM) is located between Q and plastoquinone. Studies with thylakoids isolated from atrazine-resistant pigweed plants indicate that the modification in the Q/B membrane complex that confers increased resistance to inhibition by atrazine also results in decreased sensitivity to inhibition by 2-n-heptyl-4-hydroxyquinoline-N-oxide (resistant/ sensitive ratio = 11). From the results of tetramethylphenylenediamine by-pass experiments, determinations of inhibitor sensitivity in trypsin-treated thylakoids and competitive displacement experiments made with [14C]metribuzin in thylakoids and photosystem II particles, it is suggested that 2-n-heptyl-4-hydroxyquinoline-N-oxide binds in a region of the Q/B complex that is distinct from the 3-(3,4-dichloro)-1,1-dimethyl urea and atrazine binding sites.     

Inactivation of horse liver alcohol dehydrogenase by modification of cysteine residue 174 with diazonium-1H-tetrazole.

Diazonium-1H-tetrazole was tested as a potential active-site-directed reagent for amino acid residues involved in catalysis by alcohol dehydrogenase. In a novel reaction with a protein, diazonium-1H-tetrazole inactivated the enzyme selectively, and almost stoichiometrically, but reacting with the sulfur of a cysteine residue, Cys-174. As a model compound, the tetrazole adduct of free cysteine was prepared. Elementary and spectral analyses of the adduct were consistent with the structure 5-tetrazoleazo-S-cysteine. The adduct absorbs light with a maximun at 316 nm, and is destroyed by irradiation at this wavelength. The inactivated enzyme still bound NADH as determined by difference spectroscopy, but did not enhance the fluorescence of the bound NADH as did native enzyme. X-ray crystallographic studies of free enzyme have shown that Cys-174 coordinates the zinc at the active site (Eklund, H., Nordstr?m, B., Zeppezauer, E., S?derlund, G., Ohlsson, I., Boiwe, T., and Br?ndén, C-I. (1974), FEBS Lett. 44, 200-204). The modified enzyme is probably inactive because the large, negatively charged tetrazole ring interferes sterically or electrostatically with the binding of substrates or with hydride transfer.     

1，2，4－三氯苯在土壤中的降解

在好氧和厌氧两种条件下研究了１,２,４－三氯苯的降解,结果表明,１,２,４－三氯苯的好氧降解和厌氧降解均遵循一级反应动力学在同样水分、温度及初始浓度条件下,１,２,４－三氯苯的好氧降解比厌氧降解迅速,其半衰期分别为１．８９～５．８６和５．０７～１９．０８ｄ土壤中１,２,４－三氯苯的初始浓度对于其降解也有显著影响,在０～１００μｇ·ｇ－１的范围内,浓度增高时,其降解加快,说明污染物浓度对降解的影响;在１０～３０℃范围内,温度增高导致降解过程加快,归因于温度升高对微生物酶活性的激活作用．     

Aminopyrimidines and Derivatives. 19 1. Reaction of 1, 6-Dihydro-4-Beta; D (2, 3, 4, 6-Tetra-0-Acetyl)Glucopyranosylamino-1-Methyl-Z-Methoxy-6-Oxo Pyrimidine with Choracetyl Chloride

Abstract

Reaction between 1, 6-dihydro-4-β-D(2, 3, 4, 6-tetra-O-acetyl) glucopyranosylamino-l-methyl-2-methoxy-6-oxo pyrimidine and chloracetyl chloride yields the corresponding 5-α-chloracetyl derivative and 2,6-di oxo-4-β -D (2,3,4,6-tetra-0-acetyl) glucopyranosylamino-l-methyl-1,2,3,6 tetrahdro pyrimidine. The first compund has been cyclized to the corresponding 7-β-D-glucopyranosyl-pyrrolo 2, 3-d pyrimidine and the second one to 3-βD-glucopyranosyl-vic-triazolo 4,5-d pyrimidine.