Publication Date:
2015-05-20
Description:
Phosphofructokinase-1 (PFK1), the 'gatekeeper' of glycolysis, catalyses the committed step of the glycolytic pathway by converting fructose-6-phosphate to fructose-1,6-bisphosphate. Allosteric activation and inhibition of PFK1 by over ten metabolites and in response to hormonal signalling fine-tune glycolytic flux to meet energy requirements. Mutations inhibiting PFK1 activity cause glycogen storage disease type VII, also known as Tarui disease, and mice deficient in muscle PFK1 have decreased fat stores. Additionally, PFK1 is proposed to have important roles in metabolic reprogramming in cancer. Despite its critical role in glucose flux, the biologically relevant crystal structure of the mammalian PFK1 tetramer has not been determined. Here we report the first structures of the mammalian PFK1 tetramer, for the human platelet isoform (PFKP), in complex with ATP-Mg(2+) and ADP at 3.1 and 3.4 A, respectively. The structures reveal substantial conformational changes in the enzyme upon nucleotide hydrolysis as well as a unique tetramer interface. Mutations of residues in this interface can affect tetramer formation, enzyme catalysis and regulation, indicating the functional importance of the tetramer. With altered glycolytic flux being a hallmark of cancers, these new structures allow a molecular understanding of the functional consequences of somatic PFK1 mutations identified in human cancers. We characterize three of these mutations and show they have distinct effects on allosteric regulation of PFKP activity and lactate production. The PFKP structural blueprint for somatic mutations as well as the catalytic site can guide therapeutic targeting of PFK1 activity to control dysregulated glycolysis in disease.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4510984/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4510984/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Webb, Bradley A -- Forouhar, Farhad -- Szu, Fu-En -- Seetharaman, Jayaraman -- Tong, Liang -- Barber, Diane L -- P30 DK026743/DK/NIDDK NIH HHS/ -- R01 GM047413/GM/NIGMS NIH HHS/ -- U54 GM094597/GM/NIGMS NIH HHS/ -- U54-GM094597/GM/NIGMS NIH HHS/ -- Canadian Institutes of Health Research/Canada -- England -- Nature. 2015 Jul 2;523(7558):111-4. doi: 10.1038/nature14405. Epub 2015 May 18.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Cell and Tissue Biology, University of California, San Francisco, California 94143, USA. ; Department of Biological Sciences, Northeast Structural Genomics Consortium, Columbia University, New York, New York 10027, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/25985179" target="_blank"〉PubMed〈/a〉
Keywords:
Enzyme Activation
;
Humans
;
Microscopy, Electron, Transmission
;
*Models, Molecular
;
Mutation/genetics
;
Neoplasms/*enzymology/genetics
;
Phosphofructokinase-1/*chemistry/*genetics/ultrastructure
;
Protein Structure, Tertiary
;
Recombinant Proteins/chemistry/genetics/metabolism
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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