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

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. [Display omitted] •PGAM5 catalytic domain shares phosphoglycerate mutase fold and forms stable dimer•WDPNWD motif allosterically activates the fully active dodecameric form•Crystal structures reveal conformational plasticity of the PGAM5 active site Chaikuad et al. present multiple crystal structures of PGAM5 and its biophysical analysis in solution, revealing the plasticity and structural mechanisms regulating oligomerization and its allosteric activation.