2022-12-28RELRCSBRCSBRCSB2021-10-052022-10-122022-10-122022-12-28Bill & Melinda Gates FoundationOPP1170236United StatesI53-50 nanoparticle core reconstructed from GPC-I53-50NP by focused refinementAntanasijevic ABrouwer PJMWard ABBrouwer PJMAntanasijevic ARonk AJMuller-Krauter HWatanabe YClaireaux MPerrett HRBijl TPLGrobben MUmotoy JCSchriek AIBurger JATejjani KLloyd NMSteijaert THvan Haaren MMSliepen Kde Taeye SWvan Gils MJCrispin MStrecker TBukreyev AWard ABSanders RWLassa virus glycoprotein nanoparticles elicit neutralizing antibody responses and protection.Cell Host MicrobeUS301759202236400021doi:10.1016/j.chom.2022.10.0181934-6069EMD-25108associated EM volumeEMD-25107other EM volumeEMD-25109other EM volume7sgeFULLOVERLAPI53-50 nanoparticle core recovered from GPC-I53-50 nanoparticleI53-50 nanoparticle core recovered from GPC-I53-50 nanoparticle012Nanoparticle consists of 60 monomeric I53-50A and 60 monomeric I53-50B building blocks. Nanoparticle was assembled by combining equimolar amounts of GPC-I53-50A and I53-50B.synthetic constructJosiah GPCysR4-I53-50A - nanoparticle componentsynthetic construct0.073845571Homo sapiensLEVOMGQIVTFFQEVPHVIEEVMNIVLIALSVLAVLKGLYNFATCGLVGLVTFLLLCGRSCTTSLYKGVYELQTLELNMETLNM
TMPLSCTKNNSHHYIMVGNETGLELTLTNTSIINHKFCNLSDAHKKNLYDHALMSIISTFHLSIPNFNQYEAMSCDFNGG
KISVQYNLSHSYAGDAANHCGTVANGVLQTFMRMAWGGSYIALDSGCGNWDCIMTSYQYLIIQNTTWEDHCQFSRPSPIG
YLGLLSQRTRDIYISRRRRGTFTWTLSDSEGKDTPGGYCLTRWMLIEAELKCFGNTAVAKCNEKHDEEFCDMLRLFDFNK
QAIQRLKAPAQMSIQLINKAVNALINDQLIMKNHLRDIMCIPYCNYSKYWYLNHTTTGRTSLPKCWLVSNGSYLNETHFS
DDIEQQADNMITEMLQKEYMERQGGSGGSGGSGGSGGSEKAAKAEEAARKMEELFKKHKIVAVLRANSVEEAIEKAVAVF
AGGVHLIEITFTVPDADTVIKALSVLKEKGAIIGAGTVTSVEQCRKAVESGAEFIVSPHLDEEISQFCKEKGVFYMPGVM
TPTELVKAMKLGHDILKLFPGEVVGPEFVKAMKGPFPNVKFVPTGGVDLDNVCEWFDAGVLAVGVGDALVEGDPDEVREK
AKEFVEKIRGCTEGSLEWSHPQFEKI53-50B componentsynthetic construct0.0182368771Escherichia coli BL21LEVOMNQHSHKDHETVRIAVVRARWHAEIVDACVSAFEAAMRDIGGDRFAVDVFDVPGAYEIPLHARTLAETGRYGAVLGTAFV
VNGGIYRHEFVASAVINGMMNVQLNTGVPVLSAVLTPHNYDKSKAHTLLFLALFAVKGMEAARACVEILAAREKIAAGSL
EHHHHHHsingleParticleparticle27.010.0Tris-HClTris150.0NaClSodium chlorideTBSQuantifoil R2/1COPPER400CARBONHOLEYPLASMA CLEANINGOTHERETHANE100283FEI VITROBOT MARK IVBlot time 4s, Wait time 10s, Blot force 0. I53-50 nanoparticle core recovered from GPC-I53-50 nanoparticle by focused refinement of the core. Nanoparticle was assembled by combining equimolar amounts of GPC-I53-50A and I53-50B.FEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN3002.70.61.629000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENGATAN K2 SUMMIT (4k x 4k)COUNTING2233710.550.41145508Template pickerMap reconstructed from negative stain EM1IBACK PROJECTION3.67FSC 0.143 CUT-OFFRELION3.0Icosahedral symmetry imposed for refinement. Solvent mask around the nanoparticle core used for refinement.86411OTHERRELION3.0Regularized likelihoodOTHERRELION3.0Regularized likelihoodRELION3.06P6FRIGID BODY FITREALemd_25108_msk_1.mapemd_25108_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
520
520520
0
00520520520535.6535.6535.690.090.090.0XYZ-0.00650578830.0277868943.6534915e-050.00165465111.031.031.03I53-50 nanoparticle map reconstructed from GPC-I53-50 nanoparticle by focused refinement - Half Map 2emd_25108_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
520
520520
0
00520520520535.6535.6535.690.090.090.0XYZ-0.00664179730.027524363.5950146e-050.00165399411.031.031.03I53-50 nanoparticle map reconstructed from GPC-I53-50 nanoparticle by focused refinement - Half Map 1