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Native Mass Spectrometry-Based Screening for Optimal Sample Preparation in Single-Particle Cryo-EM.
Olinares, Paul Dominic B; Kang, Jin Young; Llewellyn, Eliza; Chiu, Courtney; Chen, James; Malone, Brandon; Saecker, Ruth M; Campbell, Elizabeth A; Darst, Seth A; Chait, Brian T.
  • Olinares PDB; Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, NY 10065, USA. Electronic address: paul.olinares@rockefeller.edu.
  • Kang JY; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Llewellyn E; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Chiu C; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Chen J; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Malone B; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Saecker RM; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Campbell EA; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Darst SA; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
  • Chait BT; Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, NY 10065, USA. Electronic address: chait@rockefeller.edu.
Structure ; 29(2): 186-195.e6, 2021 02 04.
Article in English | MEDLINE | ID: covidwho-939287
Semantic information from SemMedBD (by NLM)
1. Spectrometry METHOD_OF Specimen preparation
Subject
Spectrometry
Predicate
METHOD_OF
Object
Specimen preparation
2. Single Molecule Microscopy USES Electrons
Subject
Single Molecule Microscopy
Predicate
USES
Object
Electrons
3. Spectrometry METHOD_OF Specimen preparation
Subject
Spectrometry
Predicate
METHOD_OF
Object
Specimen preparation
4. Single Molecule Microscopy USES Electrons
Subject
Single Molecule Microscopy
Predicate
USES
Object
Electrons
ABSTRACT
Recent advances in single-particle cryogenic electron microscopy (cryo-EM) have enabled the structural determination of numerous protein assemblies at high resolution, yielding unprecedented insights into their function. However, despite its extraordinary capabilities, cryo-EM remains time-consuming and resource-intensive. It is therefore beneficial to have a means for rapidly assessing and optimizing the quality of samples prior to lengthy cryo-EM analyses. To do this, we have developed a native mass spectrometry (nMS) platform that provides rapid feedback on sample quality and highly streamlined biochemical screening. Because nMS enables accurate mass analysis of protein complexes, it is well suited to routine evaluation of the composition, integrity, and homogeneity of samples prior to their plunge-freezing on EM grids. We demonstrate the utility of our nMS-based platform for facilitating cryo-EM studies using structural characterizations of exemplar bacterial transcription complexes as well as the replication-transcription assembly from the SARS-CoV-2 virus that is responsible for the COVID-19 pandemic.
Subject(s)
Keywords

Full text: Available Collection: International databases Database: MEDLINE Main subject: Mass Spectrometry / Cryoelectron Microscopy / Single Molecule Imaging Type of study: Experimental Studies Language: English Journal: Structure Journal subject: Molecular Biology / Biochemistry / Biotechnology Year: 2021 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Mass Spectrometry / Cryoelectron Microscopy / Single Molecule Imaging Type of study: Experimental Studies Language: English Journal: Structure Journal subject: Molecular Biology / Biochemistry / Biotechnology Year: 2021 Document Type: Article