PMID:15518560
| Citation |
Fenton, AW, Paricharttanakul, NM and Reinhart, GD (2004) Disentangling the web of allosteric communication in a homotetramer: heterotropic activation in phosphofructokinase from Escherichia coli. Biochemistry 43:14104-10 |
|---|---|
| Abstract |
Phosphofructokinase from Escherichia coli (EcPFK) is a homotetramer with four active sites and four allosteric sites. Understanding the allosteric activation of EcPFK by MgADP has been complicated by the complex web of possible interactions, including active site homotropic interactions, allosteric site homotropic interactions, and heterotropic interactions between active and allosteric sites. The current work has simplified this web of possible interactions to a series of single heterotropic interactions by forming and isolating hybrid tetramers. Each of the four unique heterotropic interactions have independently been isolated and compared to a control that has all four of the unique heterotropic interactions. If the interactions are labeled with the distances between interacting ligands, the 45-A interaction contributes 20% +/- 1%, the 33-A interaction contributes 34% +/- 1%, the 30-A interaction contributes 21% +/- 1%, and the 23-A interaction contributes 25% +/- 1% with respect to the total free energy of MgADP/fructose 6-phosphate (Fru-6-P) activation in the control. The free energies of the isolated interactions sum to 100% +/- 2% of the total. Therefore, the four unique interactions are all contributors to activation, are nonequivalent, and are additive. |
| Links |
PubMed Online version:10.1021/bi048569q |
| Keywords |
Adenosine Diphosphate/metabolism; Allosteric Regulation/genetics; Amino Acid Substitution/genetics; Binding Sites/genetics; Enzyme Activation/genetics; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Fructosephosphates/metabolism; Kinetics; Ligands; Mutagenesis, Site-Directed; Phosphofructokinase-1/chemistry; Phosphofructokinase-1/genetics; Phosphofructokinase-1/metabolism; Protein Subunits/chemistry; Protein Subunits/genetics; Protein Subunits/metabolism; Surface Properties |
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