Viscometric and Turbidimetric Measurements of Dilute Aqueous Solutions of a Non-ionic Detergent

Kushner, Lawrence; Hubbard, Willard D.

doi:10.1021/j150522a024

Cited by 185 publications

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“…The amount of 657 nm form of pheophytin b in a 2.3 x 101 M solution was found to be 0.76 x 10-M in 0.2% Triton X-100 and 1.4 x 10'M in 0.6% Triton X-100. Calculations of the expected rate in 0.6% Triton X-100 were made for a first order reaction and for Michaelis-Menten kinetics with the Km and VM,1 values given in Table IV Triton X-100 micelles have been estimated to have a molecular weight of 90,000 by light scattering (8,19) and 63,000 by ultracentrifugation studies (10) and to consist of 140 detergent monomers and 6000 water molecules with a radius of 43 A (19). An samples were removed at intervals for activity assays.…”

Section: Resultsmentioning

confidence: 99%

Purification and Properties of Chlorophyllase from Ailanthus altissima (Tree-of-Heaven)

1971

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Chlorophyllase from Ailanthus alissima leaves has been purified 63-fold by a combination of heat treatment, ultracentrifugation, gel filtration, and chromatography on diethylaminoethyl cellulose. While the enzyme is inhibited to some degree by Triton X-100, a modification of the assay procedure of Klein and Vishniac has been shown to be far superior to the use of aqueous acetone systems. The enzyme was found to have a pH optimum on pheophytin a of 4.5. Chlorophylls a and b, pheophytins a and b, and pyropheophytin a were hydrolyzed by the enzyme while protochlorophyll a and 4-vinyl proto. chlorophyll a were not hydrolyzed but were competitive inhibitors. p-Nitrophenyl acetate was not hydrolyzed. The enzyme does not appear to contain an essential sulfhydryl group since sodium tetrathionate and p-chloromercuribenzoate did not affect its activity.Since the discovery of chlorophyllase (chlorophyll chlorophyllidohydrolase, EC 3.1.1.14) by Willstitter and Stoll in 1913 (36), progress in our understanding of both the physiological role and of the physical, chemical, and catalytic properties of the enzyme has been quite limited. There is biological evidence consistent with both synthetic (5,7,14,15,26,30,33,34) and degradative (20,24,27,38) functions for chlorophyllase, but the data are not conclusive for either or both of these possibilities. Acquisition of definitive experimental data for this enzyme has been hindered because both the enzyme and its known substrates are insoluble in aqueous buffers. In the past decade, procedures have been developed in several laboratories (1,5,7,13,18,22,30) for solubilizing chlorophyllase from a number of plant tissues. Solubilization of substrate has been accomplished in nearly all cases by using a buffer containing from 40 to 80% acetone. Interpretation of enzyme kinetics for reactions done under these conditions is difficult. However, a more immediate problem is that substrates are only partially soluble even at high acetone concentrations (1, 3 difficult and means that quantitative assays of enzymatic activity are subject to large error. Therefore, most of the quantitative data for chlorophyllase in the literature and interpretations based upon these data are of doubtful validity.More information about the physical, chemical, and catalytic properties of chlorophyllase is needed before experiments to show the physiological function of the enzyme are likely to be successful. As a beginning to this process, we have partially purified chlorophyllase from Ailanthus altissima leaves and studied the effect of detergents, acetone concentration, pH, and ionic strength on the activity of the enzyme. Data on the effect of pH and acetone concentration on stability of the enzyme are also given. Kinetic parameters for some substrates and inhibitors of chlorophyllase have been obtained. MATERIAIS AND METHODSChlorophyllase was prepared from young leaves of wild Ailanthus altissima trees. Cellex-D (DEAE'-cellulose) from Bio-Rad Laboratories was prepared for chromatography according to th...

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Section: Resultsmentioning

confidence: 99%

Purification and Properties of Chlorophyllase from Ailanthus altissima (Tree-of-Heaven)

1971

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“…Triton treated Sie IgA-C was then titrated against untreated '25I-LDL in the 2% BSA/Tris buffer as outlined above, or untreated Sie IgA was titrated against Triton treated '251-LDL. In either case the final concentrations of Triton X-100 remained well below the critical micellar concentration for the surfactant, -0.02% (20). A parallel titration, employing nonpretreated reactants, was performed in the presence of 0.2% Triton X-100 in the assay buffer and a fourth titration was performed in the total absence of the detergent.…”

Section: Methodsmentioning

confidence: 99%

Immune complex hyperlipidemia induced by an apolipoprotein-reactive immunoglobulin A paraprotein from a patient with multiple myeloma. Characterization of this immunoglobulin.

Kilgore¹,

Patterson²,

Parenti³

et al. 1985

J. Clin. Invest.

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An antibodylike paraprotein has been isolated from a patient with multiple myeloma and autoimmune hyperlipoproteinemia.The paraprotein bound to apolipoprotein B (apo B)-containing lipoproteins that formed macromolecular aggregates, and globules thought to be aggregated complexes of lipoproteins and reactive immunoglobulins were observed circulating within the retinal blood vessels of this patient. This binding specificity permitted purification of the paraprotein from both the agglutinated immune complexes and from the plasma.The protein is an IgA, K-immunoglobulin which exists primarily in a polymeric state. Capillary immunoprecipitation demonstrated reactivity with very low density lipoproteins (VLDL) and low density proteins (LDL), but not with high density lipoproteins (HDL). Delipidated apo B and apo E, but not apo A or apo C, formed precipitates with this immunoglobulin. In using a radioimmunoassay format, the affinity of the immunoglobulin was greatest for VLDL

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“…As the concentration of the surfactant is raised through the critical micelle concentration (cmc) of about 0.25 mmol L-' (14), spherical aggregates containing approximately 140 monomers are formed (13). The diffusion coefficient of the Triton X-100 micelles, Dm is about 0.067 x m2 s-', the value determined by Weinheimer et al (15) by extrapolation to high concentrations where most of the solute is transported as micelles.…”

Section: Figmentioning

confidence: 99%

“…Other solutes, such as nucleic acids (8), amides (9-1 l), and certain dyes (12), associate in a stepwise manner to form chain-like aggregates. Non-ionic surfactants illustrate an extreme case of association in which micellar clusters containing several hundred molecules can form (13)(14)(15)(16).…”

Section: Introductionmentioning

confidence: 99%

Diffusion in associating nonelectrolyte mixtures: stepwise aggregation and micelle formation

Leaist¹

1988

Can. J. Chem.

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Equations have been developed to predict diffusion coefficients and Onsager coefficients for associating nonelectrolyte solutes in binary or multicomponent solutions with any number of association equilibria. The equations are used to interpret previously reported data for binary diffusion with stepwise association of ethanol and N-methylacetamide in carbon tetrachloride solutions. Diffusion of Triton X-100, a non-ionic micelle-forming surfactant, is also analyzed. In contrast to the gradual decrease in the diffusivity caused by stepwise association, formation of micellar aggregates produces a sharp drop in the diffusivity at the critical micelle concentration. The relation between interdiffusion coefficients and intradiffusion coefficients for associating nonelectrolyte solutes is discussed. DEREK G. LEAIST. Can. J. Chem. 66, 1129 (1988). On a dtveloppt des tquations qui permettent de prtdire les coefficients de diffusion et les coefficients dlOnsager de solutts non-tlectrolytes qui s'associent dans des solutions binaires ou B plusieurs composants et qui comportent n'importe quel nombre d'kquilibres d'association. On a utilist les tquations pour interprtter des donnkes rapporttes anttrieurement pour de la diffusion binaire impliquant une association par ttapes de l'tthanol et du N-mtthylacttamide dans des solutions de tttrachlorure de carbone. On a aussi analyst la diffusion du Triton X-100, un agent de surface non-ionique qui forme des micelles. Par opposition avec la diminution graduelle de la possibilitt de diffusion provoqute par cette association par ttapes, la formation d'aggrkgats micellaires provoque une diminution importante de la possibilitk de diffusion B la concentration micellaire critique. On discute de la relation entre les coefficients d'interdiffusion et les coefficients d'intradiffusion pour les solutks non-tlectrolytes qui s'associent.[Traduit par la revue] Introduction Many nonelectrolyte solutes associate (1). Aqueous urea, for example, is extensively dimerized (2,3). In nonpolar solvents, carboxylic acids and phenols form dimers (4-7). Other solutes, such as nucleic acids (8), amides (9-1 l), and certain dyes (12), associate in a stepwise manner to form chain-like aggregates. Non-ionic surfactants illustrate an extreme case of association in which micellar clusters containing several hundred molecules can form (13-16).The work reported here was undertaken to study the diffusion behavior of associating nonelectrolytes. The effects of association on diffusion are interpreted by determining changes in the solute mobility and the free energy gradient, the driving force for diffusion.Binary diffusion of dimerizing solutes (2,(17)(18)(19)(20) and ternary diffusion of solutes A and B that form complex AB (21) have already been studied. The equations developed in the present paper extend the earlier treatments to nonelectrolyte mixtures that contain arbitrary numbers of components and association equilibria.Weinheimer et al. (15) have reported an approximate expression for estimating the diffusion...

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Viscometric and Turbidimetric Measurements of Dilute Aqueous Solutions of a Non-ionic Detergent

Abstract: Viscometric and Turbidimetric Measurements of a Non-ionic Detergent 11G3 efficient with potassium bromide and a DVB 8 resin, and find that (Drsr) µ " is 1.

Cited by 185 publications

References 0 publications

Purification and Properties of Chlorophyllase from Ailanthus altissima (Tree-of-Heaven)

Purification and Properties of Chlorophyllase from Ailanthus altissima (Tree-of-Heaven)

Immune complex hyperlipidemia induced by an apolipoprotein-reactive immunoglobulin A paraprotein from a patient with multiple myeloma. Characterization of this immunoglobulin.

Diffusion in associating nonelectrolyte mixtures: stepwise aggregation and micelle formation

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