A glycolytic metabolon in Saccharomyces cerevisiae is stabilized by F-actin

FEBS J. 2013 Aug;280(16):3887-905. doi: 10.1111/febs.12387. Epub 2013 Jul 12.

Abstract

In the Saccharomyces cerevisiae glycolytic pathway, 11 enzymes catalyze the stepwise conversion of glucose to two molecules of ethanol plus two CO₂ molecules. In the highly crowded cytoplasm, this pathway would be very inefficient if it were dependent on substrate/enzyme diffusion. Therefore, the existence of a multi-enzymatic glycolytic complex has been suggested. This complex probably uses the cytoskeleton to stabilize the interaction of the various enzymes. Here, the role of filamentous actin (F-actin) in stabilization of a putative glycolytic metabolon is reported. Experiments were performed in isolated enzyme/actin mixtures, cytoplasmic extracts and permeabilized yeast cells. Polymerization of actin was promoted using phalloidin or inhibited using cytochalasin D or latrunculin. The polymeric filamentous F-actin, but not the monomeric globular G-actin, stabilized both the interaction of isolated glycolytic pathway enzyme mixtures and the whole fermentation pathway, leading to higher fermentation activity. The associated complexes were resistant against inhibition as a result of viscosity (promoted by the disaccharide trehalose) or inactivation (using specific enzyme antibodies). In S. cerevisiae, a glycolytic metabolon appear to assemble in association with F-actin. In this complex, fermentation activity is enhanced and enzymes are partially protected against inhibition by trehalose or by antibodies.

Keywords: actin; cytoskeleton; enzyme association; glycolytic metabolon; yeast metabolism.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / drug effects
  • Actin Cytoskeleton / metabolism*
  • Actins / agonists
  • Actins / antagonists & inhibitors
  • Actins / chemistry
  • Actins / metabolism*
  • Antibodies, Fungal / pharmacology
  • Bridged Bicyclo Compounds, Heterocyclic / pharmacology
  • Cytochalasin D / pharmacology
  • Cytoplasm / drug effects
  • Cytoplasm / enzymology
  • Cytoplasm / metabolism
  • Enzyme Stability / drug effects
  • Fermentation / drug effects
  • Glycolysis* / drug effects
  • Kinetics
  • Metabolome* / drug effects
  • Multienzyme Complexes / antagonists & inhibitors
  • Multienzyme Complexes / chemistry
  • Multienzyme Complexes / metabolism*
  • Phalloidine / pharmacology
  • Polymerization / drug effects
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / agonists
  • Saccharomyces cerevisiae Proteins / antagonists & inhibitors
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Thiazolidines / pharmacology
  • Trehalose / pharmacology
  • Tubulin Modulators / pharmacology
  • Viscosity

Substances

  • Actins
  • Antibodies, Fungal
  • Bridged Bicyclo Compounds, Heterocyclic
  • Multienzyme Complexes
  • Saccharomyces cerevisiae Proteins
  • Thiazolidines
  • Tubulin Modulators
  • Phalloidine
  • Cytochalasin D
  • Trehalose
  • latrunculin B