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TitleMolecular basis of mEAK7-mediated human V-ATPase regulation.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 3272, Year 2022
Publish dateJun 7, 2022
AuthorsRong Wang / Yu Qin / Xiao-Song Xie / Xiaochun Li /
PubMed AbstractThe activity of V-ATPase is well-known to be regulated by reversible dissociation of its V and V domains in response to growth factor stimulation, nutrient sensing, and cellular differentiation. The ...The activity of V-ATPase is well-known to be regulated by reversible dissociation of its V and V domains in response to growth factor stimulation, nutrient sensing, and cellular differentiation. The molecular basis of its regulation by an endogenous modulator without affecting V-ATPase assembly remains unclear. Here, we discover that a lysosome-anchored protein termed (mammalian Enhancer-of-Akt-1-7 (mEAK7)) binds to intact V-ATPase. We determine cryo-EM structure of human mEAK7 in complex with human V-ATPase in native lipid-containing nanodiscs. The structure reveals that the TLDc domain of mEAK7 engages with subunits A, B, and E, while its C-terminal domain binds to subunit D, presumably blocking V-V torque transmission. Our functional studies suggest that mEAK7, which may act as a V-ATPase inhibitor, does not affect the activity of V-ATPase in vitro. However, overexpression of mEAK7 in HCT116 cells that stably express subunit a4 of V-ATPase represses the phosphorylation of ribosomal protein S6. Thus, this finding suggests that mEAK7 potentially links mTOR signaling with V-ATPase activity.
External linksNat Commun / PubMed:35672408 / PubMed Central
MethodsEM (single particle)
Resolution3.73 - 4.08 Å
Structure data

26622

7une

EMDB-26622, PDB-7une:
The V1 region of bovine V-ATPase in complex with human mEAK7 (focused refinement)
Method: EM (single particle) / Resolution: 3.73 Å

26623

7unf

EMDB-26623, PDB-7unf:
CryoEM structure of a mEAK7 bound human V-ATPase complex
Method: EM (single particle) / Resolution: 4.08 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

ChemComp-POV:
(2S)-3-(hexadecanoyloxy)-2-[(9Z)-octadec-9-enoyloxy]propyl 2-(trimethylammonio)ethyl phosphate / phospholipid*YM

Source
  • bos taurus (cattle)
  • homo sapiens (human)
KeywordsPROTON TRANSPORT / mTOR signaling

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