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1.
Fungal Biol ; 124(1): 15-23, 2020 01.
Article in English | MEDLINE | ID: mdl-31892373

ABSTRACT

Metabolons are dynamic associations of enzymes catalyzing consecutive reactions within a given pathway. Association results in enzyme stabilization and increased metabolic efficiency. Metabolons may use cytoskeletal elements, membranes and membrane proteins as scaffolds. The effects of glucose withdrawal on a putative glycolytic metabolon/F-actin system were evaluated in three Saccharomyces cerevisiae strains: a WT and two different obligate fermentative (OxPhos-deficient) strains, which obtained most ATP from glycolysis. Carbon source withdrawal led to inhibition of fermentation, decrease in ATP concentration and dissociation of glycolytic enzymes from F-actin. Depending on the strain, inactivation/reactivation transitions of fermentation took place in seconds. In addition, when ATP was very low, green fluorescent protein-labeled F-actin reorganized from highly dynamic patches to large, non-motile actin bodies containing proteins and enzymes. Glucose addition restored fermentation and cytoskeleton dynamics, suggesting that in addition to ATP concentration, at least in one of the tested strains, metabolon assembly/disassembly is a factor in the control of the rate of fermentation.


Subject(s)
Actin Cytoskeleton/ultrastructure , Actins/metabolism , Cytoskeleton/enzymology , Glycolysis , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Cytoskeleton/ultrastructure , Fermentation , Glucose/metabolism , Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism , Microfilament Proteins/metabolism , Oxidative Phosphorylation , Phosphoglycerate Kinase/metabolism , Saccharomyces cerevisiae/enzymology , Saccharomyces cerevisiae/ultrastructure
2.
Mech Ageing Dev ; 87(1): 1-13, 1996 May 24.
Article in English | MEDLINE | ID: mdl-8735902

ABSTRACT

The patterns of urinary proteins in rats of different ages were examined on SDS gradient polyacrylamide gel electrophoresis coupled with silver staining. Proteins were fractionated into at least 26 bands. Densitometric measurements were used to characterize protein excretion patterns. The results showed that proteinuria in newborn, young and adult rats is predominantly tubular, consisting of low molecular-weight species. Conversely, late adults and old rats had a mixed glomerular pattern, with a steadily increasing excretion of albumin, IgG and transferrin, as was the case of other high molecular-weight proteins. Fragments of both immunoglobulins and albumin were found in all urine samples assayed. In 1 month old rats the percentage of Tamm-Hörsfall (T-H) protein was higher (P < 0.01) than in the remaining groups studied. In newborns, relatively high albumin, IgG and transferrin percentages were detected, as well as an alpha 1-acid glycoprotein and carbonic anhydrase excretion (P < 0.05 and P < 0.01 respectively) higher than that observed in the other age groups studied.


Subject(s)
Aging/urine , Proteinuria/urine , Albuminuria/urine , Animals , Animals, Newborn , Carbonic Anhydrases/urine , Electrophoresis, Polyacrylamide Gel , Immunoglobulin Fragments/urine , Male , Mucoproteins/urine , Orosomucoid/urine , Rats , Rats, Wistar , Silver , Sodium Dodecyl Sulfate , Staining and Labeling , Transferrin/urine , Uromodulin
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