Experimental basis for separation of membrane vesicles by preparative free-flow electrophoresis

Morré, D. James; Lawrence, James B.; Safranski, Keri; Hammond, Timothy G.; Morré, Dorothy M.

doi:10.1016/0021-9673(94)80110-x

Cited by 25 publications

(24 citation statements)

References 30 publications

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“…The ability of vesicles to acidify their interior in an ATPdependent process while generating a negative (outside) membrane potential might serve as a basis for enhanced electrophoretic mobility and was claimed to be an essential factor [43]. Experimental evidence was provided for ATP-induced shifts in electrophoretic mobility in membrane or organelle separation [43].…”

Section: Discussionmentioning

confidence: 98%

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Free‐flow electrophoresis for fractionation of Arabidopsis thaliana membranes

et al. 1998

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Highly purified tonoplast and plasma membrane vesicles were isolated from microsomes of Arabidopsis thaliana by preparative free-flow electrophoresis. The most electronegative fractions were identified as tonoplast using nitrate-inhibited Mg2+-ATPase as enzyme marker. The least electronegative fractions were identified as plasma membrane using glucan-synthase II, UDPG: sterol-glucosyl-transferase, and vanadate-inhibited Mg2+-ATPase as enzyme markers. Other membrane markers, latent inosine-5'-diphosphatase (Golgi), NADPH-cytochrome-c reductase (endoplasmic reticulum) and cytochrome-c oxidase (mitochondria) were recovered in the fractions intermediate between tonoplast and plasma membrane. Immunoblot analysis of membrane fractions by antibodies directed against tonoplast and plasma membrane proteins confirmed the nature and the purity of the isolated membranes. The cytoskeletal protein actin, which was also identified by immunoblotting, was found to be specifically attached to the plasma membrane vesicles. The structural and functional integrity of the isolated membranes from Arabidopsis thaliana is discussed in the light of results obtained for the location of receptors and enzymes, or for the determination of ligand binding activity.

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Section: Discussionmentioning

confidence: 98%

“…Experimental evidence was provided for ATP-induced shifts in electrophoretic mobility in membrane or organelle separation [43]. In plants, both tonoplast and plasma membrane house H+-ATPase.…”

Section: Discussionmentioning

confidence: 99%

Free‐flow electrophoresis for fractionation of Arabidopsis thaliana membranes

et al. 1998

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“…Altered proteasomal activity may alter clearance of the peptides [18]. The much larger increase in Aβ 42 than Aβ 40 suggests additional factors may be important. TD may enhance nidus and aggregation of soluble Aβ in interstitial fluids.…”

Section: Discussionmentioning

confidence: 99%

“…Critical components of protein processing occur in the endoplasmic reticulum (ER), trans-Golgi Network and Golgi apparatus where thiamine-processing enzymes also occur. Thiamine pyrophosphatase is a trans-Golgi marker enzyme [40,42]. Recent studies indicate that the thiamine dependent enzyme transketolase is localized around the endoplasmic reticulum (ER) [4].…”

Section: Discussionmentioning

confidence: 99%

Thiamine deficiency induces oxidative stress and exacerbates the plaque pathology in Alzheimer's mouse model

Karuppagounder

Shi

et al. 2009

Neurobiology of Aging

121

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Mitochondrial dysfunction, oxidative stress and reductions in thiamine-dependent enzymes have been implicated in multiple neurological disorders including Alzheimer's disease (AD). Experimental thiamine deficiency (TD) is an established model for reducing the activities of thiamine-dependent enzymes in brain. TD diminishes thiamine dependent enzymes throughout the brain, but produces a time-dependent selective neuronal loss, glial activation, inflammation, abnormalities in oxidative metabolism and clusters of degenerating neurites in only specific thalamic regions. The present studies tested how TD alters brain pathology in Tg19959 transgenic mice over expressing a double mutant form of the amyloid precursor protein (APP). TD exacerbated amyloid plaque pathology in transgenic mice and enlarged the area occupied by plaques in cortex, hippocampus and thalamus by 50%, 200% and 200%, respectively. TD increased Aβ 1-42 levels by about three-fold, β-CTF (C99) levels by 33% and β-secretase (BACE1) protein levels by 43%. TD induced inflammation in areas of plaque formation. Thus, the induction of mild impairment of oxidative metabolism, oxidative stress and inflammation induced by TD alters metabolism of APP and/or Aβ and promotes accumulation of plaques independent of neuron loss or neuritic clusters.

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“…negative outside. which mav serve as a basis Separation of DNA fragments by electrophoresis is facilitated by electrophoresis in gels or in viscous solutions of For enhanced electrophoretic mobility, was tested, e.g., with tonoplast membranes, incubated with ATP, to drive proton import or with monensin to dissipate the ATPsupported proton gradient [91]. To explore the orientation of two forms of NADH oxidase activity within the plasma membrane of HeLa cells, sealed right-side-out, plasma membranes were treated to yield a mixture of right-side-out and inside-out vesicles and the two populations were separated by preparative FFE.…”

Section: Membranes Vesicles Biological Particles and Cellsmentioning

confidence: 99%

Continuous free‐flow electrophoresis

1998

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This review evaluates the literature on continuous free flow electrophoresis, published during the last four years. Its aim is to serve not only experts in the field but also newcomers, and, therefore, it also briefly describes the principles of the method and the techniques used, referring to fundamental papers published earlier. The actual commercial instrumentation is briefly outlined. A substantial part of this review is devoted to the optimization of the performance of this method. Finally, diverse applications of fractionations of charged species in solution, ranging from small ions to biological particles and cells, are surveyed.

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Experimental basis for separation of membrane vesicles by preparative free-flow electrophoresis

Cited by 25 publications

References 30 publications

Free‐flow electrophoresis for fractionation of Arabidopsis thaliana membranes

Free‐flow electrophoresis for fractionation of Arabidopsis thaliana membranes

Thiamine deficiency induces oxidative stress and exacerbates the plaque pathology in Alzheimer's mouse model

Continuous free‐flow electrophoresis

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