Journal: Structure (London, England : 1993)

Article Title: Fusion of DARPin to aldolase enables visualization of small protein by cryoEM

doi: 10.1016/j.str.2019.04.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The sequences of the constructs tested are reported in DataS1 . table ft1 table-wrap mode="anchored" t5 caption a7 REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and Virus Strains E. coli E. cloni 10G Chemically Competent cells Lucigen Cat #60106-2 E. coli BL21(DE3) Lucigen Cat #60401-3 Deposited Data Cryo-EM map of GFP:DARPin-aldolase structure This paper EMD-9277 Coordinates of GFP:DARPin-aldolase structure This paper PDB 6MWQ Coordinates of rabbit muscle aldolase ( Herzik et al., 2017 ) PDB 5VY5 Coordinates of GFP/DARPin complex ( Hansen et al., 2017 ) PDB 5MA6 Coordinates of S. aureus protein A bound to a human IgM Fab ( Graille et al., 2000 ) PDB 1DEE Coordinates of E. coli ribosome ( Noeske et al., 2015 ) PDB 4YBB Coordinates of E. coli Beta-galactosidase ( Bartesaghi et al., 2015 ) PDB 5A1A Coordinates of V. cholerae vipA/vipB helical tube ( Kudryashev et al., 2015 ) PDB 3J9G Coordinates of EPN-01 nanocage ( Votteler et al., 2016 ) PDB 5KP9 Coordinates of E. coli TibC ( Yao et al., 2014 ) PDB 4RAP Coordinates of E. coli Ferritin ( Stillman et al., 2001 ) PBD 1EUM Recombinant DNA pACYCDuet-GFP This paper n/a pET21b-His-DARPin-aldolase This paper n/a pET21a-Beta-galactosidase-ProteinA This paper n/a pET22b-scFv This paper n/a pET21a-Beta-galactosidase-DARPin This paper n/a Software and Algorithms IMOD ( Kremer et al., 1996 ) https://bio3d.colorado.edu/imod/ Relion (Scheres, 2012b; 2012a; Zivanov et al., 2018) https://www2.mrc-lmb.cam.ac.uk/relion/index.php/Main_Page cryoSPARC ( Punjani et al., 2017 ) https://cryosparc.com/ PyMOL (DeLano, 2002) https://pymol.org/2/ COOT ( Emsley and Cowtan, 2004 ) https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ Chimera ( Pettersen et al., 2004 ) https://www.cgl.ucsf.edu/chimera/ MotionCor2 ( Zheng et al., 2017 ) http://msg.ucsf.edu/em/software/motioncor2.html CtfFind4 (Rohou and Grigorieff, 2015) http://grigoriefflab.janelia.org/ctffind4 cryoEF ( Naydenova and Russo, 2017 ) https://www.mrc-lmb.cam.ac.uk/crusso/cryoEF/ SerialEM ( Mastronarde, 2005 ) http://bio3d.colorado.edu/SerialEM/ UCSF PyEM Package n/a https://github.com/asarnow/pyem SBGrid Software Library ( Morin et al., 2013 ) https://sbgrid.org/ Other UltrAufoil® R2/2 holey gold film on Au 200 mesh grid Quantifoil, Großlöbichau, Germany Cat #687-200-AU Open in a separate window KEY RESOURCES TABLE The N- and C-termini of base proteins β-galactosidase and the EPN-01 nanocage are involved in multimerization, so the sequence of the selectable adapter was inserted within the base protein sequence.

Techniques: Virus, Ubiquitin Proteomics, Recombinant, Software