Stefania Torino / Mugdha Dhurandhar / Annelore Stroobants / Raf Claessens / Rouslan G Efremov /
PubMed Abstract
Single-particle cryogenic electron microscopy (cryo-EM) allows reconstruction of high-resolution structures of proteins in different conformations. Protein function often involves transient ...Single-particle cryogenic electron microscopy (cryo-EM) allows reconstruction of high-resolution structures of proteins in different conformations. Protein function often involves transient functional conformations, which can be resolved using time-resolved cryo-EM (trEM). In trEM, reactions are arrested after a defined delay time by rapid vitrification of protein solution on the EM grid. Despite the increasing interest in trEM among the cryo-EM community, making trEM samples with a time resolution below 100 ms remains challenging. Here we report the design and the realization of a time-resolved cryo-plunger that combines a droplet-based microfluidic mixer with a laser-induced generator of microjets that allows rapid reaction initiation and plunge-freezing of cryo-EM grids. Using this approach, a time resolution of 5 ms was achieved and the protein density map was reconstructed to a resolution of 2.1 Å. trEM experiments on GroEL:GroES chaperonin complex resolved the kinetics of the complex formation and visualized putative short-lived conformations of GroEL-ATP complex.
EMDB-16091, PDB-8bk7: Cryo-EM structure of beta-galactosidase at 3.3 A resolution plunged 5 ms after mixing with apoferritin Method: EM (single particle) / Resolution: 3.3 Å
EMDB-16092, PDB-8bk8: Cryo-EM structure of beta-galactosidase at 2.9 A resolution plunged 205 ms after mixing with apoferritin Method: EM (single particle) / Resolution: 2.9 Å
EMDB-16093, PDB-8bk9: Cryo-EM structure of mouse heavy-chain apoferritin at 2.1 A plunged 5ms after mixing with b-galactosidase Method: EM (single particle) / Resolution: 2.1 Å
EMDB-16094, PDB-8bka: Cryo-EM structure of mouse heavy-chain apoferritin at 2.7 A plunged 35ms after mixing with b-galactosidase Method: EM (single particle) / Resolution: 2.7 Å
EMDB-16095, PDB-8bkb: Cryo-EM structure of mouse heavy-chain apoferritin at 2.2 A plunged 205ms after mixing with b-galactosidase Method: EM (single particle) / Resolution: 2.2 Å
EMDB-16097, PDB-8bkg: Cryo-EM structure of beta-galactosidase at 3.2 A resolution plunged 35 ms after mixing with apoferritin Method: EM (single particle) / Resolution: 3.2 Å
EMDB-16102, PDB-8bl7: Structure of GroEL-nucleotide complex in ADP-like conformation plunged 13 ms after mixing with ATP Method: EM (single particle) / Resolution: 4.4 Å
EMDB-16108, PDB-8ble: Structure of GroEL-nucleotide complex in ADP-like conformation plunged 50 ms after mixing with ATP Method: EM (single particle) / Resolution: 4.0 Å
EMDB-16116, PDB-8bm0: Structure of GroEL:GroES-ATP complex plunge frozen 200 ms after reaction initiation Method: EM (single particle) / Resolution: 3.4 Å
EMDB-16119, PDB-8bmo: Structure of GroEL:GroES complex exhibiting ADP-conformation in trans ring obtained under the continuous turnover conditions Method: EM (single particle) / Resolution: 3.4 Å
EMDB-16125, PDB-8bmt: Structure of GroEL:GroES-ATP complex plunge frozen 200 ms after reaction initiation Method: EM (single particle) / Resolution: 2.5 Å
EMDB-16154: Map of the GroEL-ES-ATP complex plunge-frozen 50 ms after mixing with ATP Method: EM (single particle) / Resolution: 7.1 Å
EMDB-16157: Map of GroEL:GroES-(ADP) complex plunge frozen 200 ms after reaction initiation with ATP Method: EM (single particle) / Resolution: 6.3 Å
In the structure databanks used in Yorodumi, some data are registered as the other names, "COVID-19 virus" and "2019-nCoV". Here are the details of the virus and the list of structure data.
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)
EMDB accession codes are about to change! (news from PDBe EMDB page)
The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
The EM Navigator/Yorodumi systems omit the EMD- prefix.
Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator