An optimized protocol for in vitro and in cellulo structural determination of the multi-tRNA synthetase complex by cross-linking mass spectrometry

STAR Protocols

Camelia Baleanu-Gogonea¹  Krishnendu Khan²  Paul L. Fox²  Belinda Willard³  Valentin Gogonea⁴ 

[1] Corresponding author;Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44195, USA;Department of Chemistry, Cleveland State University, Cleveland, OH 44195, USA;Lerner Research Institute Proteomics and Metabolomics Core, Cleveland Clinic Foundation, Cleveland, OH 44195, USA;

关键词: Cell Biology;    Molecular Biology;    Protein Biochemistry;    Structural Biology;    Mass Spectrometry;    Chemistry;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: Despite recent advances in structural determination of individual proteins, elucidating the 3-dimensional architecture of large, multiprotein complexes remains challenging, partly because of issues related to structural integrity during purification. Here, we describe a protocol to determine the 3-dimensional architecture of the 11-constituent, multi-tRNA synthetase complex (MSC) using chemical cross-linking coupled with mass-spectrometry (XL-MS). The protocol does not require purification and is broadly applicable, facilitating determination of native structures in cell lysates and in non-disrupted cells as well as in purified complexes.For complete details on the use and execution of this protocol, please refer to Khan et al. (2020).

【 授权许可】

Unknown