The identification of the multiple components of the ROC1-cullin-based ubiquitin E3 ligase protein complex was discussed. The tryptic peptides obtained from the immunoprecipitation of Flag-ROC1 proteins were analyzed by micro-HPLZ/electropray ionization ion trap mass spectrometry (μHPLC/ESI-IT-MS) using data independent function. It was observed that low levels of signalosome/proteasome were detected in the Flag-ROC1 immunoprecipitates, which showed that the linkage between signalosome/proteasome and ROC1-cullin complexes was mediated by low affinity interactions. It was also observed that the use of ionic exchange column-based chromatography subsequent to affinity purification separated multiple ROC1-associatd protein species.
|Number of pages||2|
|State||Published - 2002|
|Event||Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics - Orlando, FL, United States|
Duration: 2 Jun 2002 → 6 Jun 2002
|Conference||Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics|
|Period||2/06/02 → 6/06/02|