Unexpected Trypsin Cleavage at Ubiquitinated Lysines

Meghan C. Burke, Yan Wang, Amanda E. Lee, Emma Kimm Dixon, Carlos A. Castaneda, David Fushman, Catherine Fenselau

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Unexpected tryptic cleavage has been characterized at modified K48 residues in polyubiquitins. In particular, the tryptic products of all seven of the lysine-linked dimers of ubiquitin and of three trimers-linear Ub-48Ub-48Ub, linear Ub-63Ub-63Ub, and the branched trimer [Ub]2-6,48Ub-have been analyzed. In addition to the peptide products expected under commonly used tryptic conditions, we observe that peptides are formed with an unexpected ε-glycinylglycinyl-Lys carboxyl terminus when the site of linkage is Lys48. Trypsin from three different commercial sources exhibited this aberration. Initial cleavage at R74 is proposed in a distal ubiquitin to produce a glycinylglycinyl-lysine residue which is bound by trypsin.

Original languageEnglish (US)
Pages (from-to)8144-8148
Number of pages5
JournalAnalytical Chemistry
Volume87
Issue number16
DOIs
StatePublished - Aug 18 2015

ASJC Scopus subject areas

  • Analytical Chemistry

Fingerprint Dive into the research topics of 'Unexpected Trypsin Cleavage at Ubiquitinated Lysines'. Together they form a unique fingerprint.

  • Cite this

    Burke, M. C., Wang, Y., Lee, A. E., Dixon, E. K., Castaneda, C. A., Fushman, D., & Fenselau, C. (2015). Unexpected Trypsin Cleavage at Ubiquitinated Lysines. Analytical Chemistry, 87(16), 8144-8148. https://doi.org/10.1021/acs.analchem.5b01960