Unexpected Trypsin Cleavage at Ubiquitinated Lysines

被引：16

作者：

Burke, Meghan C. ^{[1
]}

Wang, Yan ^{[2
]}

Lee, Amanda E. ^{[1
]}

Dixon, Emma Kimm ^{[1
]}

Castaneda, Carlos A. ^{[1
]}

Fushman, David ^{[1
]}

Fenselau, Catherine ^{[1
]}

机构：

[1] Univ Maryland, Dept Chem & Biochem, College Pk, MD 20742 USA

[2] Univ Maryland, Prote Core Facil, College Pk, MD 20742 USA

来源：

ANALYTICAL CHEMISTRY | 2015年 / 87卷 / 16期

基金：

美国国家卫生研究院;

关键词：

ACTIVE-SITE; PROTEIN; DIGESTION; SEQUENCE; STRATEGY; REVEALS; ACID;

D O I：

10.1021/acs.analchem.5b01960

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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-(48)Ub-(48)Ub, linear Ub-(63)Ub-(63)Ub, and the branched trimer [Ub](2)-(6,48)Ub-have been analyzed. In addition to the peptide products expected under commonly used tryptic conditions, we observe that peptides are formed with an unexpected epsilon-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

引用

页码：8144 / 8148

页数：5

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