Letter: Evaluation and comparison of collision-induced dissociation and electron-capture dissociation for top-down analysis of intact ribonuclease B.
It has been previously reported that the glycosylation site and protein-sequence information could be obtained for ribonuclease B by top-down electron-capture dissociation (ECD) and collision-induced dissociation (CID) mass spectrometry (MS). However, the sequence coverage of ribonuclease B was limited in a single activation, and the structural information on the glycan moiety was not probed successfully in previous experiments. Here, we demonstrate that ECD and CID techniques can be used together as an effective top- down method for the structural characterization of intact glycoprotein. Even without an elaborate pre- or post- ECD activation, a high sequence coverage (<90%) of ribonuclease B could be achieved with substantial amounts of structural information for the glycan moiety. By comparing our work with previous results, it is postulated that the disulfide bond reduction strategy might play a significant role in determining the efficiency of top-down MS.
Department of Chemistry, the Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, PR China. wangzesdas@a63.com.
Ze
Z
Wang
Department of Chemistry, the Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, PR China. Shandong Academy of Science, Shandong, 250014, PR China. xiangfchensdas@163.com.
Xiangfeng
X
Chen
Department of Chemistry, the Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, PR China. Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Washington 99352, USA.
Liulin
L
Deng
Department of Chemistry, the Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, PR China. wanlicuhk@163.com.
Wan
W
Li
Department of Chemistry, the Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, PR China. ylwongcuhk@163.com.
Y-L Elaine
YL
Wong
Department of Chemistry, the Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, PR China. twdchan@cuhk.edu.hk.
T-W Dominic
TW
Chan
D004220:Disulfides / Q000737:chemistry; D004583:Electrons; D006023:Glycoproteins / Q000737:chemistry*; D012260:Ribonucleases / Q000737:chemistry*; D020539:Sequence Analysis, Protein / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D004220:Disulfides; D006023:Glycoproteins; D012260:Ribonucleases; C029600:ribonuclease B