2024-03-06RELRCSBRCSBRCSB2020-05-202020-11-182020-11-182024-03-06Structure of human TRPA1 in complex with agonist GNE551Rohou ARouge LChen HTRPA1, channel, agonist, MEMBRANE PROTEIN, MEMBRANE PROTEIN-Agonist complexLiu CReese RVu SRouge LShields SDKakiuchi-Kiyota SChen HJohnson KShi YPChernov-Rogan TGreiner DMZKohli PBHackos DBrillantes BTam CLi TWang JSafina BMagnuson SVolgraf MPayandeh JZheng JRohou AChen JA Non-covalent Ligand Reveals Biased Agonism of the TRPA1 Ion Channel.NeuronUS109273284.e4202133152265doi:10.1016/j.neuron.2020.10.0140896-62732038NERNET6x2jFULLOVERLAPTRPA1 bound by agonist GNE551TRPA1 bound by agonist GNE55101Homo sapiensTransient receptor potential cation channel subfamily A member 1Homo sapiens0.0726221254Spodoptera frugiperdaLEVOSPLHFAASYGRINTCQRLLQDISDTRLLNEGDLHGMTPLHLAAKNGHDKVVQLLLKKGALFLSDHNGWTALHHASMGGYT
QTMKVILDTNLKCTDRLDEDGNTALHFAAREGHAKAVALLLSHNADIVLNKQQASFLHLALHNKRKEVVLTIIRSKRWDE
CLKIFSHNSPGNKCPITEMIEYLPECMKVLLDFCMLHSTEDKSCRDYYIEYNFKYLQCPLEFTKKTPTQDVIYEPLTALN
AMVQNNRIELLNHPVCKEYLLMKWLAYGFRAHMMNLGSYCLGLIPMTILVVNIKPGMAFNSTGIINETSDHSEILDTTNS
YLIKTCMILVFLSSIFGYCKEAGQIFQQKRNYFMDISNVLEWIIYTTGIIFVLPLFVEIPAHLQWQCGAIAVYFYWMNFL
LYLQRFENCGIFIVMLEVILKTLLRSTVVFIFLLLAFGLSFYILLNLQDPFSSPLLSIIQTFSMMLGDINYRESFLEPYL
RNELAHPVLSFAQLVSFTIFVPIVLMNLLIGLAVGDIADVQKHASLKRIAMQVELHTSLEKKLPLWFLRKVDQKSTIVYP
NKPRSGGMLFHIFCFLFCTGEIRQEIPNADKSLEMEILKQKYRLKDLTFLLEKQHELIKLIIQKMEIISETO757625-amino-1-[(4-bromo-2-fluorophenyl)methyl]-N-(2,5-dimethoxyphenyl)-1H-1,2,3-triazole-4-carboxamide0.0004502624ULJsingleParticleparticle0.338.19999999999999925.0Tris150.0NaClNaCl0.5TCEPQuantifoil R0.6/1GOLD300CARBONHOLEYGOLDHOLEY25.0PLASMA CLEANINGOTHER0.1Solarus plasma cleanerETHANE100277FEI VITROBOT MARK IVTriple blot.
Put blot in vitroblot
Apply 3.5ul to grid, wait 30sec, blot manually
Apply 3.5ul to grid, wait 30sec, blot manually,
apply final 3.5ul, final blot by vitrobot (3.5s). FEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN30070.02.7165000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENGIF Bioquantum20GATAN K2 QUANTUM (4k x 4k)COUNTING1-401110631.641.81505112C4FOURIER SPACE3.0FSC 0.143 CUT-OFFcisTEMData at spatial frequencies higher than 1/4.0 A-1 were not used during any part of the refinement.
The final map was density-modified using Phenix.58837RANDOM ASSIGNMENTcisTEMOTHERcisTEM5cisTEM3D reconstructionPHENIXDensity modification6PQQPDBexperimental modelAB INITIO MODELREAL