2020-11-25 REL PDBe PDBe PDBe 2019-08-30 2019-09-11 2020-01-01 2020-11-25 National Institutes of Health/National Institute Of Allergy and Infectious Diseases P50 AI150481 United States The Francis Crick Institute FC001061 United Kingdom Cryo-EM structure of human SERINC5 in LMNG micelles, bound to Fab. Rosa A Pye VE Nans A Cherepanov P Pye VE Rosa A Bertelli C Struwe WB Maslen SL Corey R Liko I Hassall M Mattiuzzo G Ballandras-Colas A Nans A Takeuchi Y Stansfeld PJ Skehel JM Robinson CV Pizzato M Cherepanov P A bipartite structural organization defines the SERINC family of HIV-1 restriction factors. Nat.Struct.Mol.Biol. US 27 78 83 2020 31907454 doi:10.1038/s41594-019-0357-0 1545-9985 EMD-10277 associated EM volume complex of SERINC5 with FAB bound to ECL4 in LMNG micelles complex of SERINC5 with FAB bound to ECL4 in LMNG micelles 0 1 2 SERINC5 Homo sapiens
Full-length, wild type human SERINC5 with C-terminal TwinStrep tag.
Homo sapiens LEVO msaqccagqlacccgsagcslccdccprirqslstrfmyalyfilvvvlccimmsttvahkmkehipffedmckgikagdtceklvgysavyrvcfgmacfffifclltlkinnskscrahihngfwffkllllgamcsgaffipdqdtflnawryvgavggflfigiqllllvefahkwnknwtagtasnklwyaslalvtlimysiatgglvlmavfytqkdscmenkillgvngglcllislvaispwvqnrqphsgllqsgviscyvtyltfsalsskpaevvldehgknvticvpdfgqdlyrdenlvtilgtslligcilyscltsttrsssdalqgryaapeleiarccfcfspggedteeqqpgkegprviydekkgtvyiysyfhfvfflaslyvmmtvtnwfnyesaniesffsgswsifwvkmascwicvllylctlvaplccptrefsvgssglevlfqgpgsggsawshpqfekgggsgggsggsawshpqfek FAB (antigen binding fragment from a monoclonal antibody) Mus musculus LEVO amino acid sequence unknown singleParticle particle 1 7.5 0.04 NaCl sodium chloride 0.01 Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid C-flat-1.2/1.3 4C GOLD 400 CARBON
No pre-treatment
 ETHANE 100 300 FEI VITROBOT MARK IV
The complex was assembled using recombinant SERINC5 in LMNG/PS and FAB and was purified by size exclusion chromatograhy immediately prior to vitrification on holey carbon EM grids.
 FEI TITAN KRIOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 300 2.7 -1.6 -4.0 FEI FALCON III (4k x 4k) COUNTING 1 7165 33.6 1 2502546 RELION 3
Ab initio reconstruction in cryoSPARC
 1 C1 FOURIER SPACE 8.2 FSC 0.143 CUT-OFF RELION 3 270151 OTHER cryoSPARC 2 OTHER cryoSPARC 2 11 RELION 3 emd_10277.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 140 140 140 0 0 0 140 140 140 305.2 305.2 305.2 90.0 90.0 90.0 X Y Z -0.06572661 0.1380292 -0.00004616876 0.005605059 2.18 2.18 2.18 0.035 AUTHOR sharpened EM map RIGID BODY FIT emd_10277_additional_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 140 140 140 0 0 0 140 140 140 305.2 305.2 305.2 90.0 90.0 90.0 X Y Z -0.020257108 0.09272604 -0.0000839550 0.004911126 2.18 2.18 2.18 AUTHOR unsharpened EM map emd_10277_half_map_2.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 140 140 140 0 0 0 140 140 140 305.2 305.2 305.2 90.0 90.0 90.0 X Y Z -0.026401546 0.10035939 -0.0000428685 0.005042684 2.18 2.18 2.18 AUTHOR EM half map 1 emd_10277_half_map_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 140 140 140 0 0 0 140 140 140 305.2 305.2 305.2 90.0 90.0 90.0 X Y Z -0.023527479 0.09883348 -0.0000494690 0.0050459313 2.18 2.18 2.18 AUTHOR EM half map 2