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The apo-structure of the leucine sensor Sestrin2 is still elusive

Journal Article · · Science Signaling
 [1];  [2];  [2];  [1]
  1. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Whitehead Inst. for Biomedical Research, Cambridge, MA (United States); Howard Hughes Medical Inst., Cambridge, MA (United States); Koch Inst. for Integrative Cancer Research, Cambridge, MA (United States); Broad Inst., Cambridge, MA (United States)
  2. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
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Sestrin2 is a GATOR2-interacting protein that directly binds leucine and is required for the inhibition of mTORC1 under leucine deprivation, indicating that it is a leucine sensor for the mTORC1 pathway. We recently reported the structure of Sestrin2 in complex with leucine [Protein Data Bank (PDB) ID, 5DJ4] and demonstrated that mutations in the leucine-binding pocket that alter the affinity of Sestrin2 for leucine result in a corresponding change in the leucine sensitivity of mTORC1 in cells. A lower resolution structure of human Sestrin2 (PDB ID, 5CUF), which was crystallized in the absence of exogenous leucine, showed Sestrin2 to be in a nearly identical conformation as the leucine-bound structure. On the basis of this observation, it has been argued that leucine binding does not affect the conformation of Sestrin2 and that Sestrin2 may not be a sensor for leucine. We show that simple analysis of the reported “apo”-Sestrin2 structure reveals the clear presence of prominent, unmodeled electron density in the leucine-binding pocket that exactly accommodates the leucine observed in the higher resolution structure. Refining the reported apo-structure with leucine eliminated the large Fobs-Fcalc difference density at this position and improved the working and free R factors of the model. Consistent with this result, our own structure of Sestrin2 crystallized in the absence of exogenous leucine also contained electron density that is best explained by leucine. Thus, the structure of apo-Sestrin2 remains elusive.
Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
National Institute of General Medical Sciences (NIGMS); National Institutes of Health (NIH); Office of Research Infrastructure Programs (ORIP); USDOD; USDOE Office of Science (SC)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1328786
Journal Information:
Science Signaling, Journal Name: Science Signaling Journal Issue: 446 Vol. 9; ISSN 1945-0877
Publisher:
AAASCopyright Statement
Country of Publication:
United States
Language:
ENGLISH

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Cited By (6)

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The Architecture of the Rag GTPase Signaling Network journal June 2017
Next Generation Strategies for Geroprotection via mTORC1 Inhibition journal February 2019
Lysosome: The metabolic signaling hub journal November 2018
Acute resistance exercise induces Sestrin2 phosphorylation and p62 dephosphorylation in human skeletal muscle journal December 2017
Nutrient sensing and TOR signaling in yeast and mammals journal December 2016

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