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Science 6 October 2006:
Vol. 314. no. 5796, pp. 109 - 112
DOI: 10.1126/science.1128868
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Reports

Extending Top-Down Mass Spectrometry to Proteins with Masses Greater Than 200 Kilodaltons

Xuemei Han,1* Mi Jin,1* Kathrin Breuker,2 Fred W. McLafferty1{dagger}

For characterization of sequence and posttranslational modifications, molecular and fragment ion mass data from ionizing and dissociating a protein in the mass spectrometer are far more specific than are masses of peptides from the protein's digestion. We extend the ~500-residue, ~50-kilodalton (kD) dissociation limitation of this top-down methodology by using electrospray additives, heated vaporization, and separate noncovalent and covalent bond dissociation. This process can cleave 287 interresidue bonds in the termini of a 1314-residue (144-kD) protein, specify previously unidentified disulfide bonds between 8 of 27 cysteines in a 1714-residue (200-kD) protein, and correct sequence predictions in two proteins, one with 2153 residues (229 kD).

1 Department of Chemistry and Chemical Biology, Baker Laboratory, Cornell University, Ithaca, NY 14853, USA.
2 Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 52a, 6020 Innsbruck, Austria.

* These authors contributed equally to this work.

{dagger} To whom correspondence should be addressed. E-mail: fwm5{at}cornell.edu

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