期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 405, 期 26, 页码 8505-8514出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-013-7267-5
关键词
Electron transfer dissociation; ETD; Electrospray; ESI; Tandem MS; MS/MS; Time-of-flight mass spectrometry; TOF MS; Top-down; Transferrin
资金
- Swiss National Science Foundation (SNF) [200021-125147/1]
Electron transfer dissociation (ETD)-based top-down mass spectrometry (MS) is the method of choice for in-depth structure characterization of large peptides, small- and medium-sized proteins, and non-covalent protein complexes. Here, we describe the performance of this approach for structural analysis of intact proteins as large as the 80 kDa serotransferrin. Current time-of-flight (TOF) MS technologies ensure adequate resolution and mass accuracy to simultaneously analyze intact 30-80 kDa protein ions and the complex mixture of their ETD product ions. Here, we show that ETD TOF MS is efficient and may provide extensive sequence information for unfolded and highly charged (around 1 charge/kDa) proteins of similar to 30 kDa and structural motifs embedded in larger proteins. Sequence regions protected by disulfide bonds within intact non-reduced proteins oftentimes remain uncharacterized due to the low efficiency of their fragmentation by ETD. For serotransferrin, reduction of S-S bonds leads to significantly varied ETD fragmentation pattern with higher sequence coverage of N- and C-terminal regions, providing a complementary structural information to top-down analysis of its oxidized form.
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