Location of Acid Phosphatase and β-Fructofuranosidase Within Yeast Cell Envelopes

Arnold, Wilfred Niels

doi:10.1128/jb.112.3.1346-1352.1972

Cited by 47 publications

(14 citation statements)

References 18 publications

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“…This enzyme is synthesized within the protoplast and then is released into a region beneath the cell wall surface, where most of it is retained in a form B essentially identical to the purified glycoprotein enzyme (for review, see reference 10). The mechanism of retention, however, has not been clear-wall-to-enzyme covalent bonding has been favored by some investigators (9, 10) and a physically restraining permeability barrier has been favored by others (1,2,8). Although not disproving the possibility of chemical bonding, our determination of a relatively fine porosity in the yeast cell wall was consistent with the idea of a permeability barrier restraining the secretion of invertase.…”

Section: Discussionsupporting

confidence: 84%

Porosity of the Yeast Cell Wall and Membrane

1974

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The limiting sizes of molecules that can permeate the intact cell wall and protoplast membrane of Saccharomyces cerevisiae were determined from the inflection points in a triphasic pattern of passive equilibrium uptake values obtained with a series of inert probing molecules varying in molecular size. In the phase identified with the yeast protoplast, the uptake-exclusion threshold corresponded to a monodisperse ethylene glycol of molecular weight = 110 and Einstein-Stokes hydrodynamic radius (rEs) = 0.42 nm. In the cell wall phase, the threshold corresponded to a polydisperse polyethylene glycol of number-average molecular weight (Mn) = 620 and average radius (rEs) = 0.81 nm. The third phase corresponded to complete exclusion of larger molecules. The assessment of cell wall porosity was confirmed by use of a second method involving analytical gel chromatographic analyses of the molecular weight distribution for a single polydisperse polyglycol before and after uptake by the cells, which indicated a quasi-monodisperse threshold for the cell wall of Mn = 760 and rEs = 0.89 nm. The results were reconciled with two situations in which much larger protein molecules previously have been reported able to penetrate the yeast cell wall.

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Section: Discussionsupporting

confidence: 84%

Porosity of the Yeast Cell Wall and Membrane

1974

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“…cell walls containing fragments of the plasma membrane) present at these high densities. The finding that most of the A6-fructofuranosidase is non-sedimentable, however, supports the suggestion of Arnold (1972) that this enzyme is located in the periplasmic space, and suggests that most of it is not covalently bound to the cell wall, as was proposed by Lampen (1968).…”

Section: Distribution (%)supporting

confidence: 85%

Distribution of membranes, especially of plasma-membrane fragments, during zonal centrifugations of homogenates from glucose-repressed Saccharomyces Cerevisiae

Nurminen

Taskinen

Suomalainen

1976

Biochemical Journal

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1. The distributions of several enzymes and other marker components were examined after zonal centrifugations of whole homogenates from glucose-repressed Saccharomyces cerevisiae on sucrose and iso-osmotic Ficoll, and the composition and morphology of the fractions were investigated. 2. After high-speed zonal centrifugation most of the protein, acid and alkaline phosphatases, alkaline pyrophosphatase, adenosine monophosphatase, beta-fructofuranosidase, alpha-mannosidase, NADPH-cytochrome c oxidoreductase and an appreciable amount of phospholipid and sterol were non-sedimentable, i.e. were at densities below 1.09 (g/cm3). Most of the RNA was at p=1.06-1.08 in Ficoll and at p=1.09-1.11 in sucrose. 3. The bulk of the Mg2+-dependent adenosine triphosphatase (Mg-ATPase) was coincident with the main peak of phospholipid and sterol, at median density 1.10, which was also rich in smooth-membrane vesicles. In Ficoll, a minor peak of phospholipid and sterol at p-1.12-1.15 contained a smaller part of the oligomycin-insensitive Mg-ATPase and heavy membrane fragments. In sucrose, several minor peaks of Mg-ATPase were in the mitochondrial density range, and a peak of oligomycin-insensitive Mg-ATPase coincident with a minor peak of phospholipid and sterol at around p-1.25 contained heavy membrane fragments of high carbohydrate content, especially mannose. 4. Further purification of the oligomycin-insensitive Mg-ATPase containing membrane preparations was performed on Urografin gradients. 5. It is argued that the oligomycin-insensitive Mg-ATPase containing membranes are fragments of the plasma membrane, but have different densities because they contain different amounts of glycoprotein particles.

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“…A 15-min pretreatment of a given yeast sample with 0.1 volume of ethylacetate (2) was taken as revealing the total enzyme concentration; degrees of expression in companion treatments are given as percentages of the total activity. Heat inactivation of 13-fructofuranosidase within standard cell suspensions of S. rouxii was conducted according to previous methodology (1).…”

Section: Methodsmentioning

confidence: 99%

“…Another unusual feature of S. rouxii is the physically cryptic nature of its P-fructofuranosidase (2, 20), an enzyme generally expressed in yeast species by dint of its location in the periplasmic space (1). Circumstantial evidence lends some support to the location of cryptic ,Bfructofuranosidase in the periplasmic bodies within young cells of S. rouxii (4,5).…”

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confidence: 99%

Effects of Polyenes, Detergents, and Other Potential Membrane Perturbants on an Osmotolerant Yeast, Saccharomyces rouxii

Arnold

Johnson

1982

Appl Environ Microbiol

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The osmotolerance of Saccharomyces rouxii 48-28 was confirmed with both NaCland KCl-fortified growth media, with more tolerance being exhibited for the potassium salt. Washed and buffered cells from unfortified medium were challenged with a variety of compounds (and also with physical treatments) that potentially would elicit membrane perturbations. The efficacy of these brief treatments was judged primarily by monitoring subsequent viability. Change in the degree of expression of P-fructofuranosidase (EC 3.2.1.26), which is cryptic in young cells of S. rouxii, was a second criterion. There was a linear correlation between cell death and enzyme expression for treatments with polyenes, detergents, some organic solvents which did not denature the enzyme, and various freeze-thaw regimens in graded amounts of glycerol. The species is relatively insensitive to polyene antimycotics, the order of decreasing effect being filipin, nystatin, and amphotericin B. S. rouxii was found to be less sensitive to osmotic shock than is Saccharomyces cerevisiae, but in neither species is P-fructofurano

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Location of Acid Phosphatase and β-Fructofuranosidase Within Yeast Cell Envelopes

Cited by 47 publications

References 18 publications

Porosity of the Yeast Cell Wall and Membrane

Porosity of the Yeast Cell Wall and Membrane

Distribution of membranes, especially of plasma-membrane fragments, during zonal centrifugations of homogenates from glucose-repressed Saccharomyces Cerevisiae

Effects of Polyenes, Detergents, and Other Potential Membrane Perturbants on an Osmotolerant Yeast, Saccharomyces rouxii

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