Charge state dependent top-down characterisation using electron transfer dissociation

Rožman, Marko; Gaskell, Simon J. (2012) Charge state dependent top-down characterisation using electron transfer dissociation. Rapid Communications in Mass Spectrometry, 26 (3). pp. 282-286. ISSN 0951-4198

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Official URL: http://onlinelibrary.wiley.com/doi/10.1002/rcm.533...

Abstract

The dissociation of protein ions (5–30 kDa) as a function of charge state has been explored in order to suggest the optimal charge state range for top-down sequencing. Proteins were generated under denaturing conditions and their charge states were modified via ion/ion proton transfer reactions prior to dissociation. Electron transfer dissociation (ETD) data suggested optimal sequence coverage for charge states in the m/z range from 700 to 950 while limited sequence coverage was noted when the precursor m/z was above 1000. Sequence coverage from ETD data was found to be dependent on protein size, with smaller proteins having better sequence coverage. An observed depletion in sequence-related information was mainly attributed to limited instrument (ion trap) performance (m/z range and resolution). For a combined ETD/collision-induced dissociation (CID) approach it is difficult to propose an optimal m/z range since good sequence coverage for CID is at intermediate charge states and the optimal m/z range increases with protein size. When only one charge state can be analysed in a combined ETD/CID approach, a range around 950 m/z is suggested as a starting point. Alternatively, two charge states should be explored, each optimal for either ETD or CID. Overall, these suggestions should be useful to achieve enhanced characterisation of smaller proteins/ large protein fragments (generated from denaturing solutions) in minimal analysis times.

Item Type:

Article

Additional Information:

"This is the peer reviewed version of the following article: Rožman, Marko; Gaskell, Simon J. (2012) Charge state dependent top-down characterisation using electron transfer dissociation. Rapid Communications in Mass Spectrometry, 26 (3). pp. 282-286, which has been published in final form http://onlinelibrary.wiley.com/doi/10.1002/rcm.5330/abstract . This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving."

Uncontrolled Keywords:

top-down; electron transfer dissociation; collision induced dissociation; protein; charge state

Subjects:

NATURAL SCIENCES > Chemistry

Divisions:

Division of Physical Chemistry

Projects: