Structural basis for allosteric regulation of human phosphofructokinase-1

Area of Science:

• Biochemistry
• Molecular Biology
• Structural Biology

Background:

• Phosphofructokinase-1 (PFK1) is crucial for glycolysis, regulating cellular energy production.
• Bacterial PFK1 regulation is well-understood as an allosteric process.
• The structural basis for eukaryotic PFK1 regulation remains largely unknown.

Purpose of the Study:

• To determine the structures of human liver PFK1 (PFKL) in active (R-state) and inactive (T-state) conformations.
• To elucidate the allosteric regulatory mechanisms of eukaryotic PFK1.
• To understand the structural basis of PFKL higher-order assembly.

Main Methods:

• Cryo-electron microscopy (cryoEM) was used to determine high-resolution structures.
• Structures of PFKL in R- and T-states were obtained.
• Structures of PFKL filaments were analyzed.

Main Results:

• The T-state structure revealed differences from bacterial PFK1 and elucidated ATP allosteric inhibition mechanisms.
• A C-terminal autoinhibitory role in stabilizing the T-state was identified.
• PFKL filament structures explained higher-order assembly, essential for cellular function.

Conclusions:

• The study provides the first structural insights into eukaryotic PFK1 allosteric regulation.
• Identified novel regulatory mechanisms and assembly principles for PFKL.
• These findings advance understanding of cellular energy metabolism control.