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A W Ruesink

Publications and source records attributed to A W Ruesink.

6 recordsLinked to original sources
PubMed1983

Orientation and integrity of plasma membrane vesicles obtained from carrot protoplasts.

Two fractions enriched in plasma membrane derived from suspension-cultured carrot (Daucus carota L.) cells were examined to determine if they differed from each other either in physical nature or in orientation. Parameters studied included the protein composition of purified membranes derived from trypsinized and nontrypsinized protoplasts as well as from trypsinized purified plasma membranes, the effect of inhibitors and membrane perturbants on ATPase activity, the binding of [acetyl-(14)C]concanavalin A to purified membrane fractions, and the competitive removal of [acetyl-(14)C]concanavalin A from purified membranes derived from [acetyl-(14)C]concanavalin A-labeled protoplasts. One fraction (at density of 1.102 grams per cubic centimeter on Renografin gradients) appears to be a mixed population of ;tightly' sealed vesicles with the majority being rightside-out vesicles of plasma membrane, and the other fraction (density 1.128 grams per cubic centimeter) apparently is a population of predominantly ;leaky' vesicles and/or nonvesicular fragments of plasma membrane, a large portion of which appear to be ;leaky' inside-out vesicles. In addition, it is shown that plasma membrane-enriched fractions can be distinguished from cellular endomembranes on the basis of protein and glycoprotein composition.

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PubMed1979

Isolation and Characterization of Concanavalin A-labeled Plasma Membranes of Carrot Protoplasts.

The plasma membranes of protoplasts released from carrot suspension culture cells were labeled with [(14)C]acetyl-concanavalin A. After homogenization a single labeled membrane fraction was isolated in a continuous isopycnic Renografin gradient. The labeled membranes peaked at an apparent density of 1.14 grams per cubic centimeter between the Golgi fraction at a density of 1.11 grams per cubic centimeter as determined by latent IDPase activity and the mitochondria at a density of 1.16 grams per cubic centimeter as determined by the cytochrome c oxidase activity. This method provided a very discrete peak of putative plasma membrane. On discontinuous Renografin gradients a relatively pure fraction of labeled plasma membranes could be readily isolated at the 1.122 to 1.146 grams per cubic centimeter interface. The labeled fraction was enriched in both an ATPase (pH 6.5) and a glucan synthetase with a pH optimum of 6.5 whose activity was promoted by magnesium and cellobiose. Enzyme activities were not altered by the membrane label.

Journal Article↗
PubMed1972

Cell Wall Regeneration around Protoplasts Isolated from Convolvulus Tissue Culture.

Protoplasts of Convolvulus arvensis L. tissue culture regenerated a wall-like structure within 3 days in culture. Although unusually electron dense and atypically amorphous in the electron microscope, this structure could be digested with Myrothecium cellulase but was resistant to protease, a Rohm and Haas pectinase, and a beta-1, 3-exoglucanase just like the original wall. A cytochemical test for callose was negative. Wall regeneration required a readily metabolized external carbon source and was not inhibited by a high concentration of cycloheximide, puromycin, or actinomycin D. Protoplast budding was correlated with the wall regeneration, and the latter was related quantitatively to the sucrose concentration in the medium. Although a concentration of 1 mum 2,4-dichlorophenoxy acetic acid is used normally for both general culture of the tissue and for wall regeneration, concentrations of 0 and 0.1 mm, which are highly deleterious to growth, have no appreciable effect on the incidence of the wall-like structure regenerated around protoplasts. The ability of protoplasts to undergo cell wall regeneration was decreased when they were cultured in the presence of proteolytic enzymes.

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PubMed1971

The plasma membrane of Avena coleoptile protoplasts.

Treatment of living protoplasts from the Avena coleoptile with enzymes and chemicals has provided new information about the external surface of the plasma membrane. Treatments with selected detergents and polyene antibiotics indicate that little sterol is present. The lysis of protoplasts in carboxymethyl-RNase which is enzymatically almost inactive provides strong evidence that the lysis previously observed in RNase is not an indication of RNA in the membrane. Divalent cations inhibit the RNase-induced lysis, indicating that such lysis involves the interaction of RNase with negatively charged sites on the plasma membrane surface. Tyrosinase treatment gives no lysis, showing that tyrosine does not play the role in these plasma membranes attributed to it in some animal cells. Peroxidase does not harm coleoptile protoplasts.

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