Quantitative structural proteomics in living cells by covalent protein painting.
Methods Enzymol
; 679: 33-63, 2023.
Article
em En
| MEDLINE
| ID: mdl-36682868
ABSTRACT
The fold and conformation of proteins are key to successful cellular function, but all techniques for protein structure determination are performed in an artificial environment with highly purified proteins. While protein conformations have been solved to atomic resolution and modern protein structure prediction tools rapidly generate near accurate models of proteins, there is an unmet need to uncover the conformations of proteins in living cells. Here, we describe Covalent Protein Painting (CPP), a simple and fast method to infer structural information on protein conformation in cells with a quantitative protein footprinting technology. CPP monitors the conformational landscape of the 3D proteome in cells with high sensitivity and throughput. A key advantage of CPP is its' ability to quantitatively compare the 3D proteomes between different experimental conditions and to discover significant changes in the protein conformations. We detail how to perform a successful CPP experiment, the factors to consider before performing the experiment, and how to interpret the results.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteoma
/
Proteômica
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
Methods Enzymol
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Estados Unidos