The Glycolytic Enzyme Phosphofructokinase-1 Assembles into Filaments

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2017 Jun 25

PMID 28646105

Citations 58

Authors

Bradley A Webb

Anne M Dosey

Torsten Wittmann

Justin M Kollman

Diane L Barber

Affiliations

Soon will be listed here.

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.

Citing Articles

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References

Albe K, Butler M, Wright B . Cellular concentrations of enzymes and their substrates. J Theor Biol. 1990; 143(2):163-95. DOI: 10.1016/s0022-5193(05)80266-8. View

Rohou A, Grigorieff N . CTFFIND4: Fast and accurate defocus estimation from electron micrographs. J Struct Biol. 2015; 192(2):216-21. PMC: 6760662. DOI: 10.1016/j.jsb.2015.08.008. View

Sievers F, Wilm A, Dineen D, Gibson T, Karplus K, Li W . Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol. 2011; 7:539. PMC: 3261699. DOI: 10.1038/msb.2011.75. View

Cohen R, JEDZINIAK J, Benedek G . The functional relationship between polymerization and catalytic activity of beef liver glutamate dehydrogenase. II. Experiment. J Mol Biol. 1976; 108(1):179-99. DOI: 10.1016/s0022-2836(76)80102-7. View

Trujillo J, Deal Jr W . Pig liver phosphofructokinase: asymmetry properties, proof of rapid association--dissociation equilibria, and effect of temperature and protein concentration on the equilibria. Biochemistry. 1977; 16(14):3098-104. DOI: 10.1021/bi00633a009. View

Wang J, Morris A, Tolan D, Pagliaro L . The molecular nature of the F-actin binding activity of aldolase revealed with site-directed mutants. J Biol Chem. 1996; 271(12):6861-5. View

De Bock K, Georgiadou M, Schoors S, Kuchnio A, Wong B, Cantelmo A . Role of PFKFB3-driven glycolysis in vessel sprouting. Cell. 2013; 154(3):651-63. DOI: 10.1016/j.cell.2013.06.037. View

Segall J, Tyerech S, Boselli L, Masseling S, Helft J, Chan A . EGF stimulates lamellipod extension in metastatic mammary adenocarcinoma cells by an actin-dependent mechanism. Clin Exp Metastasis. 1996; 14(1):61-72. DOI: 10.1007/BF00157687. View

Jang S, Nelson J, Bend E, Rodriguez-Laureano L, Tueros F, Cartagenova L . Glycolytic Enzymes Localize to Synapses under Energy Stress to Support Synaptic Function. Neuron. 2016; 90(2):278-91. PMC: 4840048. DOI: 10.1016/j.neuron.2016.03.011. View

10.

Reinhart G, LARDY H . Rat liver phosphofructokinase: use of fluorescence polarization to study aggregation at low protein concentration. Biochemistry. 1980; 19(7):1484-90. DOI: 10.1021/bi00548a035. View

11.

Scheres S . RELION: implementation of a Bayesian approach to cryo-EM structure determination. J Struct Biol. 2012; 180(3):519-30. PMC: 3690530. DOI: 10.1016/j.jsb.2012.09.006. View

12.

Beckner M, Stracke M, Liotta L, Schiffmann E . Glycolysis as primary energy source in tumor cell chemotaxis. J Natl Cancer Inst. 1990; 82(23):1836-40. DOI: 10.1093/jnci/82.23.1836. View

13.

Li P, Banjade S, Cheng H, Kim S, Chen B, Guo L . Phase transitions in the assembly of multivalent signalling proteins. Nature. 2012; 483(7389):336-40. PMC: 3343696. DOI: 10.1038/nature10879. View

14.

Barry R, Bitbol A, Lorestani A, Charles E, Habrian C, Hansen J . Large-scale filament formation inhibits the activity of CTP synthetase. Elife. 2014; 3:e03638. PMC: 4126345. DOI: 10.7554/eLife.03638. View

15.

Shaikh T, Gao H, Baxter W, Asturias F, Boisset N, Leith A . SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs. Nat Protoc. 2009; 3(12):1941-74. PMC: 2737740. DOI: 10.1038/nprot.2008.156. View

16.

Egelman E . Reconstruction of helical filaments and tubes. Methods Enzymol. 2010; 482:167-83. PMC: 3245864. DOI: 10.1016/S0076-6879(10)82006-3. View

17.

Ingerson-Mahar M, Briegel A, Werner J, Jensen G, Gitai Z . The metabolic enzyme CTP synthase forms cytoskeletal filaments. Nat Cell Biol. 2010; 12(8):739-46. PMC: 3210567. DOI: 10.1038/ncb2087. View

18.

KEMP R . Rabbit liver phosphofructokinase. Comparison of some properties with those of muscle phosphofructokinase. J Biol Chem. 1971; 246(1):245-52. View

19.

OConnell J, Zhao A, Ellington A, Marcotte E . Dynamic reorganization of metabolic enzymes into intracellular bodies. Annu Rev Cell Dev Biol. 2012; 28:89-111. PMC: 4089986. DOI: 10.1146/annurev-cellbio-101011-155841. View

20.

Webb B, Forouhar F, Szu F, Seetharaman J, Tong L, Barber D . Structures of human phosphofructokinase-1 and atomic basis of cancer-associated mutations. Nature. 2015; 523(7558):111-4. PMC: 4510984. DOI: 10.1038/nature14405. View