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Structures of PGAM5 Provide Insight into Active Site Plasticity and Multimeric Assembly

DOI: 10.1016/j.str.2017.05.020 DOI Help

Authors: Apirat Chaikuad (Institute for Pharmaceutical Chemistry, Johann Wolfgang Goethe-University and Buchmann Institute for Molecular Life Sciences; Structural Genomics Consortium and Target Discovery Institute, University of Oxford) , Panagis Filippakopoulos (Structural Genomics Consortium and Target Discovery Institute, University of Oxford; Ludwig Institute for Cancer Research, University of Oxford) , Sean R. Marcsisin (Northeastern University) , Sarah Picaud (Structural Genomics Consortium and Target Discovery Institute, University of Oxford; Ludwig Institute for Cancer Research, University of Oxford) , Martin Schroeder (Institute for Pharmaceutical Chemistry, Johann Wolfgang Goethe-University and Buchmann Institute for Molecular Life Sciences) , Shiori Sekine (The University of Tokyo) , Hidenori Ichijo (The University of Tokyo) , John R. Engen (Northeastern University) , Kohsuke Takeda (Nagasaki University) , Stefan Knapp (Institute for Pharmaceutical Chemistry, Johann Wolfgang Goethe-University and Buchmann Institute for Molecular Life Sciences; Structural Genomics Consortium and Target Discovery Institute, University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Structure

State: Published (Approved)
Published: June 2017
Diamond Proposal Number(s): 6391

Open Access Open Access

Abstract: PGAM5 is a mitochondrial membrane protein that functions as an atypical Ser/Thr phosphatase and is a regulator of oxidative stress response, necroptosis, and autophagy. Here we present several crystal structures of PGAM5 including the activating N-terminal regulatory sequences, providing a model for structural plasticity, dimerization of the catalytic domain, and the assembly into an enzymatically active dodecameric form. Oligomeric states observed in structures were supported by hydrogen exchange mass spectrometry, size-exclusion chromatography, and analytical ultracentrifugation experiments in solution. We report that the catalytically important N-terminal WDPNWD motif acts as a structural integrator assembling PGAM5 into a dodecamer, allosterically activating the phosphatase by promoting an ordering of the catalytic loop. Additionally the observed active site plasticity enabled visualization of essential conformational rearrangements of catalytic elements. The comprehensive biophysical characterization offers detailed structural models of this key mitochondrial phosphatase that has been associated with the development of diverse diseases.

Journal Keywords: phosphoglycerate mutase; PGAM5; Ser/Thr phosphatase; catalysis; oligomerization; allosteric regulation; WDXNWD motif; histidine acid phosphatase; active site plasticity

Subject Areas: Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography

Documents:
1-s2.0-S0969212617301545-main.pdf