The uneven distribution of Mg2+, K+, and phosphate in Saccharomyces carlsbergensis was demonstrated by the differential extraction ions. Their concentrations were 5, 60, 1 mM cytoplasm 73, 470, 110 vacuoles, respectively. intracellular gradients these ions 1:15, 1:8, 1:110, respectively, across tonoplast. determination free Mg2+ (1.35 cytosol 20 vacuoles) showed that ion accumulation vacuoles could not be explained higher degree complexing organelles.

Manganese transport into yeast cells is energy-dependent. It dependent on endogenous sources of energy and inhibited by olygomycin (12.5–25 μg/ml), 2,4-dinitrophenol (1 mM), 2-deoxyglucose (1–50 mM) sodium azide (1–10 but stimulated cyanide glucose. The stimulating effect glucose eliminated N-ethylmaleimide iodoacetate, which apparently inhibit the itself. About 75% manganese accumulated in presence found protoplasts nearly 25% cell walls. A major portion vacuoles. concentration osmotically...

Abstract Saccharomyces cerevisiae cell envelope polyphosphatase was isolated in highly active and stable form by extraction from cells with zwittergent TM‐314 followed chromatography of the extract on phosphocellulose QAE‐Sephadex presence 5 m M ‐MgCl 2 , 0·5 ‐EDTA 0·1% Triton X‐100. The enzyme possessed a specific activity 220 U/mg after 30 days retained 87% its at −20°C. Polyphosphatase molecular mass determined to be about 40 kDa gel filtration polyacrylamide electrophoresis sodium...

The secondary transport systems of the yeast vacuolar membrane have been investigated by (i) method radioactive isotopes ([ 14 C]arginine); (ii) activation H + ‐ATPase cations (Cat ), when enzyme is under control and (iii) measurement changes in proton gradient (Δ pH) potential ( E m ) due to supposed substrates transporters. main mechanism cation across tonoplast probably /Cat antiport. apparent K antiporters for Ca 2+ , Mg Mn Zn P i are 0.06, 0.3, 0.8, 0.055‐0.17 1.5mM, respectively.

Saccharomyces carlsbergensis cells accumulated Mn2+ (or Mg2+) ions in the presence of glucose, fructose, or mannose, but not deoxyglucose, 3-O-methylglucose, and sorbose. Accumulation one equivalent Mn/2+ was coupled with efflux two equivalents K+ from cells. Mg/2+ did exit during uptake. Preliminary treatment various proton conductors glucose led to loss proportional inhibition Polyene antibiotic candicidin together elicited rapid completely inhibited accumulation. Exogenous (more than 1...

The 450-million-year-old interaction between the majority of land plants and mycorrhizal fungi is one most ancient, abundant, ecologically important symbiosis on earth. early events in evolution symbioses seem to have involved reciprocal genetic changes ancestral free-living fungi. new data mechanism action specific signaling molecules how it influence influenced by membrane ions fluxes cytoplasm ion oscillations which integrate symbiotic ionome are improving our understanding molecular...

CaThi is a thionin-like peptide isolated from fruits of Capsicum annuum, which has strong antimicrobial activity against bacteria, yeasts and filamentous fungi, induced reactive oxygen species (ROS) in fungi. ROS are molecules that appear the early stages programmed cell death or apoptosis Due to this fact, work we analyzed some events may be related process on yeast by CaThi. To investigate possibility, evaluated phosphatidylserine (PS) externalization, presence active caspases ability bind...

Here we describe the phenotypic characterization of cta4+ gene, encoding a novel member P4 family P-type ATPases fission yeast. The cta4Δ mutant is temperature sensitive and cold lethal displays several morphological defects in cell polarity cytokinesis. Microtubules are generally destabilized cells lacking Cta4p. microtubule length decreased, number microtubules per increased. This concomitant with an increase catastrophe events midzone cell. These likely due to general imbalance cation...

The effect of n-alcohols on ATP-dependent generation delta pH and Em across the plasma membrane vesicles yeast Saccharomyces carlsbergensis was investigated. alcohols were shown to collapse in order C2 less than C3 C4 C5 or equal C6 greater C7 C8 C11, dissipation being more pronounced. Inhibition plasmalemma H(+)-ATPase insignificant; at low alcohol concentrations its activity even increased. basic reason for toxic cells suggested be due increase anion proton permeability membrane. Mg2+...

Vacuoles of yeast grown in peptone medium possessed high ATPase activity (up to 1 mumol X mg protein-1 min-1). Membrane-bound and solubilized activities were insensitive vanadate azide, but inhibited by NO-3 . K+ cyclic AMP stimulated both membrane-bound activities. Dio-9 activated the membrane form vacuolar 1.5-2-fold did not affect enzyme. Solubilized partially purified was reconstituted with soy-bean phospholipids a freeze-thaw procedure. native vacuoles proteoliposomes effectively Dio-9,...

This study establishes the role of P(5A)-type Cta4 ATPase in Ca(2+) sequestration endoplasmic reticulum by detecting an ATP-dependent, vanadate-sensitive and FCCP insensitive (45)Ca(2+)-transport fission yeast membranes isolated cellular fractionation. Specifically, Ca(2+)-ATPase transport activity was decreased ER from cells lacking a cta4(+) gene. Furthermore, disruption resulted 6-fold increase intracellular levels, sensitivity towards accumulation misfolded proteins stress, stimulation...

The PMR1 gene of Saccharomyces cerevisiae is thought to encode a putative Ca2+-ATPase [1]. Membranes isolated from wild-type cells and pmr1 null mutant S. were fractionated on sucrose density gradients. In the we found decrease in activity P-type ATPase ATP-dependent, protonophore-insensitive Ca2+ transport light membranes, that comigrate with Golgi marker GDPase. We conclude product (Pmr1p) indeed Golgi-like membranes. Surprisingly, mutation abolished and/or membranes only 50% under...

Lipid transfer proteins (LTP) facilitate of lipids between membranes in vitro. Up to now, they have been found be localized basically the plant cell wall and compartments linked lipid metabolism, such as glyoxysomes. Accordingly, LTP are considered involved defence against pathogen microbes metabolism. We herein show, by immunoelectron microscopy, that besides wall, lumen organelles which we suggest protein storage vacuoles, well vesicles similar lipid‐containing ones extracellular space...

Polyacrylamide gel electrophoresis (PAGE) of partially purified ATPase from vacuoles Saccharomyces carlsbergensis under non-dissociating conditions revealed 3 bands with activity. Further PAGE in dissociating showed the similarity composition these preparations. They were assumed to contain same vacuolar exhibiting, however, different electrophoretic mobility due formation enzyme complexes proteins and phospholipids. The preparation highest contained 6 subunits 75, 62, 16, 14, 12 9 kDa....

Polyamines play a regulatory role in eukaryotic cell growth and morphogenesis. Despite many molecular advances, the underlying mechanism of action remains unclear. Here, we investigate by which spermine affects morphogenesis dimorphic fungal model emerging relevance plant interactions, Yarrowia lipolytica, through recruitment phytohormone-like pathway involving an activation plasma membrane P-type H+-ATPase. Morphological transition was followed microscopically H+-ATPase activity analyzed...

A novel primary Ca 2+ ‐transport system in membrances from Saccharomyces cerevisiae is described. transport strictly dependent on the presence of ATP; other nucleotides like GTP, UTP and CTP do not efficiently (<10% rate ATP) drive uptake. Transport inhibited by sodium vanadate with an IC 50 130 μM, but insensitive to carbonylcyanide p ‐trifluoromethoxy‐phenylhydrazone, valinomycin, gramicidin or calmodulin. accumulates a free form can be readily released ionophore A‐23187 osmotic shock....