A Variety of Activation Methods Employed in ¡°Activated-Ion¡± Electron Capture Dissociation Mass Spectrometry: A Test against Bovine Ubiquitin 7+ Ions

Bulletin of the Korean chemical society

Fred W. McLafferty¹  HanBin Oh¹ 

关键词: Electron capture dissociation (ECD);    Fourier-transform mass spectrometry (FTMS);    Ubiquitin;    Activated-ion ECD;    In-beam ECD;   

DOI  :  

学科分类：化学（综合）

来源: Korean Chemical Society

【 摘 要 】

Fragmentation efficiencies of various ‘activated-ion�? electron capture dissociation (AI-ECD) methods are compared for a model system of bovine ubiquitin 7+ cations. In AI-ECD studies, sufficient internal energy was given to protein cations prior to ECD application using IR laser radiation, collisions, blackbody radiation, or in-beam collisions, in turn. The added energy was utilized in increasing the population of the precursor ions with less intra-molecular noncovalent bonds or enhancing thermal fluctuations of the protein cations. Removal of noncovalent bonds resulted in extended structures, which are ECD friendly. Under their best conditions, a variety of activation methods showed a similar effectiveness in ECD fragmentation. In terms of the number of fragmented inter-residue bonds, IR laser/blackbody infrared radiation and ‘in-beam�? activation were almost equally efficient with ~70% sequence coverage, while collisions were less productive. In particular, ‘in-beam�? activation showed an excellent effectiveness in characterizing a pre-fractionated single kind of protein species. However, its inherent procedure did not allow for isolation of the protein cations of interest.

【 授权许可】

Unknown