Fusion of isolated biological membranes

Gratzl, Manfred; Ekerdt, Roland; Schudt, Christian; Dahl, Gerhard

doi:10.5282/ubm/epub.7492

1980

DOI: 10.5282/ubm/epub.7492

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Fusion of isolated biological membranes

Manfred Gratzl¹,

Roland Ekerdt²,

Christian Schudt³

et al.

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1983

1986

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Association of Intracellular Low-density Vesicles with Plasma Membranes from Saccharomyces cerevisiae NCYC 366

et al. 1983

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Fractionation on sucrose density gradients of spheroplast lysates from Saccharomyces cerevisiae NCYC 366 yielded a fraction with a peak density of 1-05 g ml-l, intermediate between that of membranes and intracellular low-density vesicles. Electron microscopy showed the fraction to consist of membranes associated with intracellular vesicles. Evidence for the presence of plasma membranes in the association fraction was obtained by using 2sI-labelled spheroplasts. The fraction was free from detectable amounts of cytochromes and DNA. Fractionation on sucrose density gradients of incubation mixtures containing isolated crude plasma-membranes and vesicles gave rise to a visible intermediate-density band, which electron microscopy showed to contain membranes associated with vesicles. Evidence for the presence of plasma membranes in the in vitro intermediate-density band came from incubating mixtures containing * I-labelled crude plasma-membranes, and evidence for the presence of vesicles came from using 251-labelled vesicles. Supplementing incubation mixtures with calcium chloride sharply increased the size of the intermediate-density band. Cycloheximide and methylbenzimidazol-2-ylcarbamate had no effect on its formation. Purified plasma-membranes failed to form an intermediate-density band when incubated with vesicles. Supplementing these incubation mixtures with calcium chloride did not produce an intermediate-density band, but caused extensive association of vesicles with plasma membranes that pelleted in gradients. Isolated vesicles were not osmotically active ; their polyphosphate content was 1.61 pmol orthophosphate equivalent (mg vesicle protein)-'. They had diameters in the range 0-45-0-65 pm, as measured on electron micrographs of thin sections. The data reported provide further evidence for a role for intracellular low-density vesicles in envelope growth in S. cereuisiae.

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Association of Intracellular Low-density Vesicles with Plasma Membranes from Saccharomyces cerevisiae NCYC 366

et al. 1983

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Oosporogenesis by Lagenidium giganteum: induction and maturation are regulated by calcium and calmodulin

Kerwin¹,

Washino²

1986

Can. J. Microbiol.

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Induction and maturation of the sexual stage (oospores) of the facultative mosquito parasite Lagenidium giganteum (Oomycetes: Lagenidiales) are complex developmental processes regulated by calcium-dependent events. Use of developmentally synchronized cultures of L. giganteum allowed stage-specific disruption of calcium metabolism. A calcium chelator (EGTA), an ionophore (chlortetracycline), and inhibitors of the calcium-binding protein calmodulin (dibucaine, trifluoperazine, chlorpromazine) disrupted several discrete developmental steps associated with oosporogenesis: induction of antheridia, gametangial fusion, meiosis, oospore wall formation, and subsequent spore maturation. Extracellular calcium is necessary for oosporogenesis to proceed normally and under some conditions magnesium has a synergistic effect with calcium on oospore induction. Results are discussed in relation to calcium mediation of fusion events in a number of model membrane and biological systems.

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Fusion of isolated biological membranes

Cited by 2 publications

References 0 publications

Association of Intracellular Low-density Vesicles with Plasma Membranes from Saccharomyces cerevisiae NCYC 366

Association of Intracellular Low-density Vesicles with Plasma Membranes from Saccharomyces cerevisiae NCYC 366

Oosporogenesis by Lagenidium giganteum: induction and maturation are regulated by calcium and calmodulin

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