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991.
Renografin density gradient centrifugation and carboxymethyl (CM)-and diethylaminoethyl (DEAE)-cellulose ion-exchange chromatography were compared for purification of cells of phony peach, rickettsia-like bacterium (RLB) from plant extracts. Centrifugation on 30–40% linear gradients resulted in four bands. Cells of RLB sedimented into the third band located 4.9–5.4 cm below the meniscus. Phase-contrast microscopy, plant peroxidase activity, and viable bacterial assays showed the resulting cells ofRLB to be relatively free of host contaminants. A typical purification from 120 ml of extract containing 7.2×109
RLB cells resulted in the recovery of 7.0×107
RLB cells. Cells ofRLB were not successfully purified byCM andDEAE chromatography. Unitil a medium is available for cultivation of plantRLB, Renografin density gradient centrifugation should be a useful tool for obtaining cells ofRLB for biochemical and serological investigations. 相似文献
992.
Nine styrylpyridine analogs were tested as inhibitors of choline acetyltransferase which had been highly purified from rat cerebrum and bovine caudate nuclei. In general, concentrations required to achieve 50% inhibiion (I50 values) were in the micromolar range. For some analogs, I50 values were similar to those obtained previously by other investigators who used less purified enzyme preparations. With certain analogs, however, the measured values of I50 changed as the transferase became more purified, which may indicate the presence in the extract of other molecules which can interact with the enzyme. The methods used in purification of the enzyme suggest that the molecule which modifies the activity of CAT is probably a protein. The mode of inhibition by naphthylvinylpyridinium was found to be uncompetitive with respect to both choline and acetyl coenzyme A for both the rat and bovine transferases. 相似文献
993.
Daniel P. Rossignol Aimee J. Roschelle William J. Lennarz 《Journal of cellular biochemistry》1981,15(4):347-358
We have attempted to identify a surface component of echinoderm eggs that is involved in the species-specific binding of sperm. Cell surface membranes from eggs of the sea urchins Strongylocentrotus purpuratus or Arbacia punctulata were radioiodinated, detergent-treated, and subjected to density-gradient centrifugation. In the presence of bindin, the complementary binding protein isolated from sperm, one component of the membranes sedimented to a different density. This membrane component bound-species specifically to sperm that had undergone the acrosome reaction. This binding led to an inhibition of the ability of treated sperm to fertilize eggs. Exhaustive proteolytic digestion of this receptor fraction yields a high molecular weight glycopeptide that can also bind to bindin. It therefore appears that this egg surface membrane fraction contains a functionally intact, species-specific receptor for sperm. 相似文献
994.
Sea urchin sperm–egg adhesion is mediated by bindin, a sperm surface protein that has lectin-like activity. Bindin agglutinates eggs, and this interaction has been shown to be inhibited by glycopeptides released from the egg surface by protease treatment. In this study, we report the purification and properties of such an egg surface glycoconjugate that may be involved in sperm adhesion. The glycoconjugate was partially purified by gel filtration and affinity chromatography on bindin particles. Upon gel filtration on Sepharose CL 4-B, the glycoconjugate elutes near the void volume, suggesting that it has a molecular weight in excess of one million. In addition, we have found that the egg surface glycoconjugate agglutinates bindin particles, indicating that it is multivalent. Carbohydrate analysis indicates that the glycoconjugate is composed primarily of fucosc, xylose, galactose, and glucose. This purified egg surface component is the most potent inhibitor of bindin-mediated egg agglutination yet described. 相似文献
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William R. Tolbert Joseph Peder Richard C. Kimes 《In vitro cellular & developmental biology. Plant》1981,17(10):885-890
Summary A system has been developed for growth and maintenance of mammalian cells in suspension culture at high density. In principle,
the maintenance of constant levels of required nutrients coupled with the removal of toxic cell byproducts can support much
higher suspension cell densities than may be obtained in conventional spinners. The system consisted of 4- or 40-liter reaction
vessels equipped with a vertically supported rotating cylindrical filter. Agitation was provided by the magnetically driven,
rotating filter. Fresh medium was supplied at a rate of 10 to 20 ml/h per 109 cells and the expended medium free of cells was withdrawn through the rotating filter. Both pH and dissolved O2 and CO2 were monitored and regulated. Walker 256 carcinosarcoma cells have been grown in these reactors to densities 10-to 30-fold
greater than that obtained in Bellco spinners. In addition to high cell densities, the yield of cells per liter of medium
used was 2- to 3-fold that obtained in the conventional systems. Both 4-and 40-liter versions of this reactor have been operated
without the use of antibiotics. The 40-liter reactor also has been modified for chemostat operation. In a single run, for
example, the Walker cell density was maintained between 6 and 10×106 cells/ml with a total yield of 8.7×1011 cells from 360 liters of medium. 相似文献