The allosteric regulation of pyruvate kinase by fructose-1,6-bisphosphate
Open accessAbstract
BACKGROUND: Yeast pyruvate kinase (PK) catalyzes the final step in glycolysis. The enzyme therefore represents an important control point and is allosterically activated by fructose-1,6-bisphosphate (FBP). In mammals the enzyme is found as four different isozymes with different regulatory properties: two of these isozymes are produced by alternate splicing. The allosteric regulation of PK is directly related to proliferation of certain cell types, as demonstrated by the expression of an allosterically regulated isozyme in tumor cells. A model for the allosteric transition from the inactive (T) state to the active (R) state has been proposed previously, but until now the FBP-binding site had not been identified. RESULTS: We report here the structures of PK from yeast complexed with a substrate analog and catalytic metal ions in the presence and absence of bound FBP. The allosteric site is located 40 A from the active site and is entirely located in the enzyme regulatory (C) domain. A phosphate-binding site for the allosteric activator is created by residues encoded by a region of the gene corresponding to the alternately spliced exon of mammalian isozymes. FBP activation appears to induce several conformational changes among active-site sidechains through a mechanism that is most likely to involve significant domain motions, as previously hypothesized. CONCLUSIONS: The structure and location of the allosteric activator site agrees with the pattern of alternate genetic splicing of the PK gene in multicellular eukaryotes that distinguishes between a non-regulated isozyme and the regulated fetal isozymes. The conformational differences observed between the active sites of inactive and fully active PK enzymes is in agreement with the recently determined thermodynamic mechanism of allosteric activation through a 'metal relay' that increases the affinity of the enzyme for its natural phosphoenolpyruvate substrate.
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The pyruvate kinase activity of PKM2 tetramers can be regulated by the concentration of FBP, serine, and by post-translational modifications including tyrosine phosphorylation.
“The pyruvate kinase activity of the tetramers can be regulated by the concentration of the naturally occurring upstream glycolytic intermediate FBP (Ashizawa et al., 1991; Jurica et al., 1998), serine (Chaneton et al., 2012), as well as by a multitude of post-translational modifications (Wu and Le, 2013) including tyrosine phosphorylation.”
From Pyruvate kinase M2 regulates Hif-1α activity and IL-1β induction and is a critical determinant of the Warburg effect in LPS-activated macrophages (2015) · cites this paper as Jurica et al., 1998
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- Authors
- Jurica, M. S., Mesecar, A., Heath, P. J., Shi, W., Nowak, T., & Stoddard, B. L.
- Affiliations
- University of Washington (US)
- University of California, Berkeley (US)
- Fred Hutch Cancer Center (US)
- Pennsylvania State University (US)
- University of Notre Dame (US)
- Corresponding author
- Stoddard, Barry L
- Venue
- Structure
- Publisher
- Elsevier BV
- Published
- 1998-02
- Volume
- 6
- Issue
- 2
- Pages
- 195-210
- ISSN
- 0969-2126 (ISSN-L)
- OpenAlex
- W2066969376
- Cited by
- 426
- References
- 71
- Citations by year
- 2026: 8 · 2025: 19 · 2024: 16 · 2023: 16 · 2022: 22 · 2021: 29 · 2020: 21 · 2019: 35 · 2018: 26 · 2017: 29 · 2016: 24 · 2015: 17 · 2014: 18 · 2013: 18 · 2012: 20
- FWCI
- 5.14
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Structure (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- National Science Foundation
- National Institutes of Health
- Topics
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
- Keywords
- pyruvate kinase, fructose-1,6-bisphosphate, allosteric regulation, glycolysis, alternative splicing, pyruvate kinase isozymes, conformational change, catalytic metal ions, phosphate binding site
- MeSH terms
- Allosteric Regulation, Amino Acid Sequence, Binding Sites, Fructosediphosphates, Genes, Glycolates, Manganese, Models, Molecular, Molecular Sequence Data, Phosphoenolpyruvate, Potassium, Protein Conformation, Pyruvate Kinase, Saccharomyces cerevisiae, Alternative Splicing
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