Intracellular NADPH levels affect the oligomeric state of the glucose 6-phosphate dehydrogenase.
In the yeast Kluyveromyces lactis, glucose 6-phosphate dehydrogenase (G6PDH) is detected as two differently migrating forms on native polyacrylamide gels. The pivotal metabolic role of G6PDH in K. lactis led us to investigate the mechanism controlling the two activities in respiratory and fermentative mutant strains. An extensive analysis of these mutants showed that the NAD(+)(H)/NADP(+)(H)-dependent cytosolic alcohol (ADH) and aldehyde (ALD) dehydrogenase balance affects the expression of the G6PDH activity pattern. Under fermentative/ethanol growth conditions, the concomitant activation of ADH and ALD activities led to cytosolic accumulation of NADPH, triggering an alteration in the oligomeric state of the G6PDH caused by displacement/release of the structural NADP(+) bound to each subunit of the enzyme. The new oligomeric G6PDH form with faster-migrating properties increases as a consequence of intracellular redox unbalance/NADPH accumulation, which inhibits G6PDH activity in vivo. The appearance of a new G6PDH-specific activity band, following incubation of Saccharomyces cerevisiae and human cellular extracts with NADP(+), also suggests that a regulatory mechanism of this activity through NADPH accumulation is highly conserved among eukaryotes.
Dipartimento di Biologia e Biotecnologia C. Darwin, Sapienza Università di Roma, Rome, Italy. michele.saliola@uniroma1.it
Michele
M
Saliola
Angela
A
Tramonti
Claudio
C
Lanini
Samantha
S
Cialfi
Daniela
D
De Biase
Claudio
C
Falcone
D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007716:Kluyveromyces / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009249:NADP / Q000378:metabolism*; D010084:Oxidation-Reduction; D055503:Protein Multimerization; D021122:Protein Subunits / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D021122:Protein Subunits; D009249:NADP; D000426:Alcohol Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D000444:Aldehyde Dehydrogenase