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Información de la secuencia primaria de las proteínas intactas por electrospray ionización espectrometría de masas en
J A Loo1, C G Edmonds, R D Smith
1Chemical Sciences Department, Pacific Northwest Laboratory, Richland, WA 99352.
Science (New York, N.Y.)
|April 13, 1990
Resumen
La espectrometría de masas en tándem (MS/MS) analizó la ribonucleasa A bovina intacta, revelando información de la secuencia primaria. La estructura de la proteína influye en los patrones de fragmentación durante el análisis.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- Química Analítica La Química Analítica es la
- La proteómica es la proteómica.
Sus antecedentes:
- La espectrometría de masas en tándem (MS/MS) es una poderosa técnica para el análisis de proteínas.
- La ionización por electrospray (ESI, por sus siglas en inglés) genera iones de proteína cargados para la EM/EM.
- Comprender la estructura de las proteínas es crucial para interpretar los datos de la espectrometría de masas.
Objetivo del estudio:
- Para obtener información de la secuencia primaria de la ribonucleasa A bovina intacta utilizando MS/MS.
- Investigar la influencia de la estructura de proteínas de orden superior en los patrones de fragmentación.
Principales métodos:
- La ribonucleasa A bovina intacta fue analizada utilizando ionización por electrospray y espectrometría de masas en tándem (MS/MS) en un instrumento triple cuádrupolo.
- Se empleó la disociación inducida por colisión (CID, por sus siglas en inglés) para fragmentar iones moleculares cargados múltiples.
- Se compararon los espectros de EM/EM tanto de la ribonucleasa A nativa como de la reducida por disulfuro.
Principales resultados:
- El análisis de MS/MS arrojó fragmentos de iones que pudieron ser asignados a la secuencia primaria conocida de la ribonucleasa A.
- La disociación de iones de carga múltiple produjo iones de producto complementarios.
- Se observaron diferencias en los espectros de EM/EM entre las formas nativas y las reducidas en disulfuro, lo que indica una influencia estructural.
Conclusiones:
- La espectrometría de masas en tándem puede proporcionar información de secuencia primaria para proteínas intactas.
- La estructura de la proteína de la fase gaseosa afecta significativamente las vías de disociación y los patrones de fragmentación en la EM/EM.
- Este enfoque ofrece información sobre las relaciones estructura-función de las proteínas.
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