2024-03-20RELRCSBRCSBRCSB2018-12-062019-03-132019-03-132024-03-20National Institutes of Health/National Human Genome Research Institute (NIH/NHGRI)DP2EB020402United StatesSingle-particle cryo-EM reconstruction of human methemoglobin using 200 keV, state 1Herzik Jr MAWu Mheme-binding, hetero-4-mer, globin, OXYGEN TRANSPORTHerzik Jr MAWu MLander GCHigh-resolution structure determination of sub-100 kDa complexes using conventional cryo-EM.Nat CommunUK1010321032201930833564doi:10.1038/s41467-019-08991-82041-17236nbcFULLOVERLAPmethemoglobin from humanmethemoglobin from human012Lyophilized human methemoglobin purchased from Sigma AldrichHomo sapiens0.064Hemoglobin subunit alphaHomo sapiens0.0149931589999999992LEVOVLSPADKTNVKAAWGKVGAHAGEYGAEALERMFLSFPTTKTYFPHFDLSHGSAQVKGHGKKVADALTNAVAHVDDMPNAL
SALSDLHAHKLRVDPVNFKLLSHCLLVTLAAHLPAEFTPAVHASLDKFLASVSTVLTSKYP69905Hemoglobin subunit betaHomo sapiens0.0154596972LEVOVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLGAFSDGLAHLDN
LKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVANALAHP68871PROTOPORPHYRIN IX CONTAINING FE0.0006164874HEMsingleParticleparticle127.510.0Na2HPO4Phosphate137.0NaClSodium chloride2.7KCLPotassium Chloride2.0KH2PO4Potassium PhosphateQuantifoil, UltrAuFoil, R1.2/1.3GOLD300PLASMA CLEANINGOTHERGrids were plasma cleaned using a Solarus plasma cleaner (Gatan, Inc.).ETHANE90277.15HOMEMADE PLUNGERSample was manually blotted for 4-5 seconds using Whatman No. 1 filter paper immediately prior to plunge-freezing.. Lyophilized human methemoglobin (Sigma Aldrich) was solubilized.FEI TALOS ARCTICAFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN20070.02.75.016.073000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENGATAN K2 SUMMIT (4k x 4k)COUNTING371038381-442167311.069.01Counting mode, 250 ms frames, exposure rate of ~1.95 e- pixel-1 s-1, total exposure of ~69 e- angstrom-2 (1.57 e- angstrom-2 frame-1).1232543Difference-of-Gaussian-picked.An ab initio model was generated from 160,169 template-picked particles using cryoSPARC and low-pass filtered to 20 A for use as an initial model for 3D refinement.C22.8FSC 0.143 CUT-OFFRELION2.124308MAXIMUM LIKELIHOODRELION2.1MAXIMUM LIKELIHOODRELION2.14N7PAPDBexperimental modelFLEXIBLE FITREAL67.0emd_0407_msk_1.mapemd_0407_additional_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
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00256256256142.848142.848142.84890.090.090.0XYZ-0.0118884890.039866690.000145158690.00175233290.5580.5580.558Single-particle cryo-EM reconstruction of methemoglobin state 1 (unsharpened)emd_0407_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
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00256256256142.848142.848142.84890.090.090.0XYZ-0.032640720.050515660.000130670870.003470760.5580.5580.558Odd half mapemd_0407_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)