Impact of Chemical Cross-Linking on Protein Structure and Function.
Anal Chem
; 90(2): 1104-1113, 2018 01 16.
Article
en En
| MEDLINE
| ID: mdl-29232109
Chemical cross-linking coupled with mass spectrometry is a popular technique for deriving structural information on proteins and protein complexes. Also, cross-linking has become a powerful tool for stabilizing macromolecular complexes for single-particle cryo-electron microscopy. However, an effect of cross-linking on protein structure and function should not be forgotten, and surprisingly, it has not been investigated in detail so far. Here, we used kinetic studies, mass spectrometry, and NMR spectroscopy to systematically investigate an impact of cross-linking on structure and function of human carbonic anhydrase and alcohol dehydrogenase 1 from Saccharomyces cerevisiae. We found that cross-linking induces rather local structural disturbances and the overall fold is preserved even at a higher cross-linker concentration. The results establish general experimental conditions for chemical cross-linking with minimal effect on protein structure and function.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Alcohol Deshidrogenasa
/
Anhidrasas Carbónicas
/
Reactivos de Enlaces Cruzados
Límite:
Humans
Idioma:
En
Revista:
Anal Chem
Año:
2018
Tipo del documento:
Article
País de afiliación:
República Checa
Pais de publicación:
Estados Unidos