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Structure paper

TitleStructures and gating mechanisms of human bestrophin anion channels.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 3836, Year 2022
Publish dateJul 4, 2022
AuthorsAaron P Owji / Jiali Wang / Alec Kittredge / Zada Clark / Yu Zhang / Wayne A Hendrickson / Tingting Yang /
PubMed AbstractBestrophin-1 (Best1) and bestrophin-2 (Best2) are two members of the bestrophin family of calcium (Ca)-activated chloride (Cl) channels with critical involvement in ocular physiology and direct ...Bestrophin-1 (Best1) and bestrophin-2 (Best2) are two members of the bestrophin family of calcium (Ca)-activated chloride (Cl) channels with critical involvement in ocular physiology and direct pathological relevance. Here, we report cryo-EM structures of wild-type human Best1 and Best2 in various states at up to 1.8 Å resolution. Ca-bound Best1 structures illustrate partially open conformations at the two Ca-dependent gates of the channels, in contrast to the fully open conformations observed in Ca-bound Best2, which is in accord with the significantly smaller currents conducted by Best1 in electrophysiological recordings. Comparison of the closed and open states reveals a C-terminal auto-inhibitory segment (AS), which constricts the channel concentrically by wrapping around the channel periphery in an inter-protomer manner and must be released to allow channel opening. Our results demonstrate that removing the AS from Best1 and Best2 results in truncation mutants with similar activities, while swapping the AS between Best1 and Best2 results in chimeric mutants with swapped activities, underlying a key role of the AS in determining paralog specificity among bestrophins.
External linksNat Commun / PubMed:35789156 / PubMed Central
MethodsEM (single particle)
Resolution1.78 - 3.11 Å
Structure data

EMDB-27127, PDB-8d1e:
hBest2 1uM Ca2+ (Ca2+-bound) closed state
Method: EM (single particle) / Resolution: 1.78 Å

EMDB-27128, PDB-8d1f:
hBest2 5mM Ca2+ (Ca2+-bound) closed state
Method: EM (single particle) / Resolution: 1.82 Å

EMDB-27129, PDB-8d1g:
hBest2 Ca2+-bound open state
Method: EM (single particle) / Resolution: 2.07 Å

EMDB-27130, PDB-8d1h:
hBest2 Ca2+-unbound closed state
Method: EM (single particle) / Resolution: 1.94 Å

EMDB-27131, PDB-8d1i:
hBest1 1uM Ca2+ (Ca2+-bound) closed state
Method: EM (single particle) / Resolution: 1.82 Å

EMDB-27132, PDB-8d1j:
hBest1 5mM Ca2+ (Ca2+-bound) closed state
Method: EM (single particle) / Resolution: 2.05 Å

EMDB-27133, PDB-8d1k:
hBest1 Ca2+-bound partially open neck state
Method: EM (single particle) / Resolution: 2.28 Å

EMDB-27134, PDB-8d1l:
hBest1 Ca2+-bound partially open aperture state
Method: EM (single particle) / Resolution: 2.12 Å

EMDB-27135, PDB-8d1m:
hBest1 Ca2+-unbound closed state
Method: EM (single particle) / Resolution: 3.11 Å

EMDB-27136, PDB-8d1n:
bBest2_345 Ca2+-bound open state
Method: EM (single particle) / Resolution: 1.93 Å

EMDB-27137, PDB-8d1o:
hBest1_345 Ca2+-bound open state
Method: EM (single particle) / Resolution: 2.44 Å

Chemicals

ChemComp-CA:
Unknown entry

ChemComp-MC3:
1,2-DIMYRISTOYL-RAC-GLYCERO-3-PHOSPHOCHOLINE / phospholipid*YM

ChemComp-DU0:
2-[2-[(1~{S},2~{S},4~{S},5'~{R},6~{R},7~{S},8~{R},9~{S},12~{S},13~{R},16~{S})-5',7,9,13-tetramethylspiro[5-oxapentacyclo[10.8.0.0^{2,9}.0^{4,8}.0^{13,18}]icos-18-ene-6,2'-oxane]-16-yl]oxyethyl]propane-1,3-diol

ChemComp-CL:
Unknown entry

ChemComp-HOH:
WATER

Source
  • homo sapiens (human)
  • bos taurus (cattle)
KeywordsTRANSPORT PROTEIN / Ion channel / chloride channel / anion channel / pentamer

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