Structural Investigation of a Dimeric Variant of Pyruvate Kinase Muscle Isoform 2
Abstract
Pyruvate kinase muscle isoform 2 (PKM2) catalyzes the terminal step in glycolysis, transferring a phosphoryl group from phosphoenolpyruvate to ADP, to produce pyruvate and ATP. PKM2 activity is allosterically regulated by fructose 1,6-bisphosphate (FBP), an upstream glycolytic intermediate. FBP stabilizes the tetrameric form of the enzyme. In its absence, the PKM2 tetramers dissociate, yielding a dimer–monomer mixture having lower enzymatic activity. Here, the S437Y variant of PKM2 is incapable of binding FBP. Consistent with that defect, we find that S437Y exists in a monomer–dimer equilibrium in solution, with a Kd of ~20 μM. Interestingly, however, the protein crystallizes as a tetramer, providing insight into the structural basis for impaired FBP binding of S437Y.
- Authors:
-
[1]
- Univ. of Iowa, Iowa City, IA (United States)
- Univ. of Missouri—Columbia, MO (United States)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES); National Science Foundation (NSF)
- OSTI Identifier:
- 1423366
- Grant/Contract Number:
- AC02-05CH11231; CLP 1506181
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biochemistry
- Additional Journal Information:
- Journal Volume: 56; Journal Issue: 50; Journal ID: ISSN 0006-2960
- Publisher:
- American Chemical Society (ACS)
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; peptides and proteins; monomers; crystal structure; oligomers; chemical structure; pyruvate kinase; glycolysis; size-exclusion chromatography; small angle X-ray scattering; analytical ultracentrifuge
Citation Formats
Srivastava, Dhiraj, Razzaghi, Mortezaali, Henzl, Michael T., and Dey, Mishtu. Structural Investigation of a Dimeric Variant of Pyruvate Kinase Muscle Isoform 2. United States: N. p., 2017.
Web. doi:10.1021/acs.biochem.7b01013.
Srivastava, Dhiraj, Razzaghi, Mortezaali, Henzl, Michael T., & Dey, Mishtu. Structural Investigation of a Dimeric Variant of Pyruvate Kinase Muscle Isoform 2. United States. https://doi.org/10.1021/acs.biochem.7b01013
Srivastava, Dhiraj, Razzaghi, Mortezaali, Henzl, Michael T., and Dey, Mishtu. Tue .
"Structural Investigation of a Dimeric Variant of Pyruvate Kinase Muscle Isoform 2". United States. https://doi.org/10.1021/acs.biochem.7b01013. https://www.osti.gov/servlets/purl/1423366.
@article{osti_1423366,
title = {Structural Investigation of a Dimeric Variant of Pyruvate Kinase Muscle Isoform 2},
author = {Srivastava, Dhiraj and Razzaghi, Mortezaali and Henzl, Michael T. and Dey, Mishtu},
abstractNote = {Pyruvate kinase muscle isoform 2 (PKM2) catalyzes the terminal step in glycolysis, transferring a phosphoryl group from phosphoenolpyruvate to ADP, to produce pyruvate and ATP. PKM2 activity is allosterically regulated by fructose 1,6-bisphosphate (FBP), an upstream glycolytic intermediate. FBP stabilizes the tetrameric form of the enzyme. In its absence, the PKM2 tetramers dissociate, yielding a dimer–monomer mixture having lower enzymatic activity. Here, the S437Y variant of PKM2 is incapable of binding FBP. Consistent with that defect, we find that S437Y exists in a monomer–dimer equilibrium in solution, with a Kd of ~20 μM. Interestingly, however, the protein crystallizes as a tetramer, providing insight into the structural basis for impaired FBP binding of S437Y.},
doi = {10.1021/acs.biochem.7b01013},
journal = {Biochemistry},
number = 50,
volume = 56,
place = {United States},
year = {Tue Nov 28 00:00:00 EST 2017},
month = {Tue Nov 28 00:00:00 EST 2017}
}
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Works referencing / citing this record:
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