Isolation and characterization of nuclear envelope fragments from <i>dictyostelium</i>

Widmer, Rosa M.; Parish, Roger W.

doi:10.1016/0014-5793(80)81293-2

Cited by 7 publications

(1 citation statement)

References 33 publications

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“…The size and morphology of the ER membranes containing ProtA-NuoL N were studied by cryo electron microscopy (Frederik and Hubert 2005). High salt concentrations during the isolation of the ER membranes were avoided because they are known to result in extensive breakdown of the nuclear envelope (Agutter 1972;Bornens and Courvalin 1978;Widmer and Parish 1980). Figure 2 shows the micrographs of ER membranes with and without ProtA-NuoL N embedded in vitrous ice.…”

Section: Resultsmentioning

confidence: 99%

Transport of Na+ and K+ by an antiporter-related subunit from the Escherichia coli NADH dehydrogenase I produced in Saccharomyces cerevisiae

et al. 2007

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The NADH dehydrogenase I from Escherichia coli is a bacterial homolog of the mitochondrial complex I which translocates Na + rather than H + . To elucidate the mechanism of Na + transport, the C-terminally truncated NuoL subunit (NuoL N ) which is related to Na + /H + antiporters was expressed as a protein A fusion protein (ProtANuoL N ) in the yeast Saccharomyces cerevisiae which lacks an endogenous complex I. The fusion protein inserted into membranes from the endoplasmatic reticulum (ER), as conWrmed by diVerential centrifugation and Western analysis. Membrane vesicles containing ProtA-NuoL N catalyzed the uptake of Na + and K + at rates which were signiWcantly higher than uptake by the control vesicles under identical conditions, demonstrating that ProtA-NuoL N translocated Na + and K + independently from other complex I subunits. Na + transport by ProtA-NuoL N was inhibited by EIPA (5-(N-ethyl-N-isopropyl)-amiloride) which speciWcally reacts with Na + /H + antiporters. The cation selectivity and function of the NuoL subunit as a transporter module of the NADH dehydrogenase complex is discussed.

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