Interaction between phosphofructokinase and aldolase from Saccharomyces cerevisiae studied by aqueous two-phase partitioning

Phosphofructokinase (EC 2.7.1.11) and aldolase (EC 4.1.2.13) have been high ly purified from Saccharomyces cerevisiae by improved protocols. Partitioni ng of the enzymes in aqueous polymer two-phase systems was used to detect c omplex formation. The partition of each enzyme was found to be affected by the presence of the other enzyme. AMP affected the partition of the individ ual enzymes as well as the mixture of the two. The activities of the respec tive enzymes were stimulated in the putative complex in an AMP-dependent ma nner. Two strictly conserved residues belonging to an acidic surface loop o f class II aldolases, are a potential site for electrostatic interaction wi th the positively charged regions close to the active site in phosphofructo kinase. (C) 2001 Elsevier Science B.V. All rights reserved.