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TitleStructural basis of closed groove scrambling by a TMEM16 protein.
Journal, issue, pagesNat Struct Mol Biol, Vol. 31, Issue 10, Page 1468-1481, Year 2024
Publish dateApr 29, 2024
AuthorsZhang Feng / Omar E Alvarenga / Alessio Accardi /
PubMed AbstractActivation of Ca-dependent TMEM16 scramblases induces phosphatidylserine externalization, a key step in multiple signaling processes. Current models suggest that the TMEM16s scramble lipids by ...Activation of Ca-dependent TMEM16 scramblases induces phosphatidylserine externalization, a key step in multiple signaling processes. Current models suggest that the TMEM16s scramble lipids by deforming the membrane near a hydrophilic groove and that Ca dependence arises from the different association of lipids with an open or closed groove. However, the molecular rearrangements underlying groove opening and how lipids reorganize outside the closed groove remain unknown. Here we directly visualize how lipids associate at the closed groove of Ca-bound fungal nhTMEM16 in nanodiscs using cryo-EM. Functional experiments pinpoint lipid-protein interaction sites critical for closed groove scrambling. Structural and functional analyses suggest groove opening entails the sequential appearance of two π-helical turns in the groove-lining TM6 helix and identify critical rearrangements. Finally, we show that the choice of scaffold protein and lipids affects the conformations of nhTMEM16 and their distribution, highlighting a key role of these factors in cryo-EM structure determination.
External linksNat Struct Mol Biol / PubMed:38684930
MethodsEM (single particle)
Resolution2.64 - 4.07 Å
Structure data

EMDB-41453, PDB-8toi:
nhTMEM16 lipid scramblase in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (closed state)
Method: EM (single particle) / Resolution: 2.64 Å

EMDB-41454, PDB-8tok:
nhTMEM16 lipid scramblase in lipid nanodiscs with MSP2N2 scaffold protein in the presence of Ca2+ (open state)
Method: EM (single particle) / Resolution: 3.84 Å

EMDB-41455, PDB-8tol:
nhTMEM16 lipid scramblase in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (open state)
Method: EM (single particle) / Resolution: 2.85 Å

EMDB-41457: nhTMEM16 lipid scramblase in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (closed state) (consensus map)
Method: EM (single particle) / Resolution: 2.64 Å

EMDB-41458: nhTMEM16 lipid scramblase in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (closed state)(local refined monomer map)
Method: EM (single particle) / Resolution: 2.64 Å

EMDB-41477, PDB-8tpm:
nhTMEM16 lipid scramblase in lipid nanodiscs with MSP1E3 scaffold protein in the absence of Ca2+
Method: EM (single particle) / Resolution: 2.93 Å

EMDB-41478, PDB-8tpn:
nhTMEM16 lipid scramblase in lipid nanodiscs with MSP2N2 scaffold protein in the presence of Ca2+ (intermediate state)
Method: EM (single particle) / Resolution: 4.07 Å

EMDB-41479, PDB-8tpo:
nhTMEM16 R432A mutant in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+
Method: EM (single particle) / Resolution: 3.59 Å

EMDB-41480, PDB-8tpp:
nhTMEM16 R432A mutant in lipid nanodiscs with MSP2N2 scaffold protein in the presence of Ca2+
Method: EM (single particle) / Resolution: 3.64 Å

EMDB-41481, PDB-8tpq:
nhTMEM16 A444P mutant in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (long TM6)
Method: EM (single particle) / Resolution: 3.35 Å

EMDB-41482, PDB-8tpr:
nhTMEM16 A444P mutant in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (short TM6)
Method: EM (single particle) / Resolution: 2.85 Å

EMDB-41483, PDB-8tps:
nhTMEM16 A444P mutant in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (bent TM6)
Method: EM (single particle) / Resolution: 2.97 Å

EMDB-41484, PDB-8tpt:
nhTMEM16 A444P mutant in lipid nanodiscs with MSP1E3 scaffold protein in the presence of Ca2+ (long TM6/short TM6)
Method: EM (single particle) / Resolution: 3.01 Å

Chemicals

ChemComp-CA:
Unknown entry

ChemComp-PGW:
(1R)-2-{[(S)-{[(2S)-2,3-dihydroxypropyl]oxy}(hydroxy)phosphoryl]oxy}-1-[(hexadecanoyloxy)methyl]ethyl / phospholipid*YM

Source
  • fusarium vanettenii 77-13-4 (fungus)
  • fusarium vanettenii (fungus)
KeywordsLIPID TRANSPORT / membrane protein / lipid scramblase / TMEM16

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