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J Cell Biol. 2017 Aug 7;216(8):2305-2313. doi: 10.1083/jcb.201701084. Epub 2017 Jun 23.

The glycolytic enzyme phosphofructokinase-1 assembles into filaments.

Author information

1
Department of Cell and Tissue Biology, University of California, San Francisco, San Francisco, CA.
2
Department of Biochemistry, University of Washington, Seattle, WA.
3
Department of Cell and Tissue Biology, University of California, San Francisco, San Francisco, CA diane.barber@ucsf.edu.

Abstract

Despite abundant knowledge of the regulation and biochemistry of glycolytic enzymes, we have limited understanding on how they are spatially organized in the cell. Emerging evidence indicates that nonglycolytic metabolic enzymes regulating diverse pathways can assemble into polymers. We now show tetramer- and substrate-dependent filament assembly by phosphofructokinase-1 (PFK1), which is considered the "gatekeeper" of glycolysis because it catalyzes the step committing glucose to breakdown. Recombinant liver PFK1 (PFKL) isoform, but not platelet PFK1 (PFKP) or muscle PFK1 (PFKM) isoforms, assembles into filaments. Negative-stain electron micrographs reveal that filaments are apolar and made of stacked tetramers oriented with exposed catalytic sites positioned along the edge of the polymer. Electron micrographs and biochemical data with a PFKL/PFKP chimera indicate that the PFKL regulatory domain mediates filament assembly. Quantified live-cell imaging shows dynamic properties of localized PFKL puncta that are enriched at the plasma membrane. These findings reveal a new behavior of a key glycolytic enzyme with insights on spatial organization and isoform-specific glucose metabolism in cells.

PMID:
28646105
PMCID:
PMC5551713
DOI:
10.1083/jcb.201701084
[Indexed for MEDLINE]
Free PMC Article

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