Mass spectrometry guided in situ proteolysis to obtain crystals for X-ray structure determination.
J Am Soc Mass Spectrom
; 21(10): 1795-801, 2010 Oct.
Article
en En
| MEDLINE
| ID: mdl-20685133
ABSTRACT
A strategy for increasing the efficiency of protein crystallization/structure determination with mass spectrometry has been developed. This approach combines insights from limited proteolysis/mass spectrometry and crystallization via in situ proteolysis. The procedure seeks to identify protease-resistant polypeptide chain segments from purified proteins on the time-scale of crystal formation, and subsequently crystallizing the target protein in the presence of the optimal protease at the right relative concentration. We report our experience with 10 proteins of unknown structure, two of which yielded high-resolution X-ray structures. The advantage of this approach comes from its ability to select only those structure determination candidates that are likely to benefit from application of in situ proteolysis, using conditions most likely to result in formation of a stable proteolytic digestion product suitable for crystallization.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Conformación Proteica
/
Espectrometría de Masas
/
Proteínas
Idioma:
En
Revista:
J Am Soc Mass Spectrom
Año:
2010
Tipo del documento:
Article
País de afiliación:
Estados Unidos