Coupled regulations of enzymatic activity and structure formation of aldehyde dehydrogenase Ald4p

Biol Open. 2020 Apr 28;9(4):bio051110. doi: 10.1242/bio.051110.


Previously, we have developed an extramitochondrial assembly system, where mitochondrial targeting signal (MTS) can be removed from a given mitochondrial enzyme, which could be used to characterize the regulatory factors involved in enzyme assembly/disassembly in vivo Here, we demonstrate that addition of exogenous acetaldehyde can quickly induce the supramolecular assembly of MTS-deleted aldehyde dehydrogenase Ald4p in yeast cytoplasm. Also, by using PCR-based modification of the yeast genome, cytoplasmically targeted Ald4p cannot polymerize into long filaments when key functional amino acid residues are substituted, as shown by N192D, S269A, E290K and C324A mutations. This study has confirmed that extramitochondrial assembly could be a powerful external system for studying mitochondrial enzyme assembly, and its regulatory factors outside the mitochondria. In addition, we propose that mitochondrial enzyme assembly/disassembly is coupled to the regulation of a given mitochondrial enzyme activity.

Keywords: Aldehyde dehydrogenase; Extramitochondrial assembly; Yeast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Dehydrogenase / chemistry*
  • Aldehyde Dehydrogenase / genetics
  • Aldehyde Dehydrogenase / metabolism*
  • Amino Acid Sequence
  • Binding Sites
  • Catalytic Domain
  • Enzyme Activation
  • Fluorescent Antibody Technique
  • Mitochondria / enzymology
  • Models, Molecular*
  • Protein Binding
  • Protein Conformation*
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Structure-Activity Relationship


  • Recombinant Fusion Proteins
  • Aldehyde Dehydrogenase