Screening ubiquitin specific protease activities using chemically synthesized ubiquitin and ubiquitinated peptides

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• 作者：Marine Bacchi^a ; Benjamin Fould^a ; Magali Jullian^b ; Aude Kreiter^b ; Amé ; lie Maurras^b ; Olivier Nosjean^a ; Thibault Coursindel^b ; Karine Puget^b ; Gilles Ferry^a ; Jean A. Boutin^a ; ^{jean.boutin@servier.com}
• 关键词：Ubiquitinated peptides ; Chemical synthesis ; Deubiquitinase assay ; HPLC
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：15 February 2017
• 年：2017
• 卷：519
• 期：Complete
• 页码：57-70
• 全文大小：2675 K
• 卷排序：519

文摘

Chemically synthesized ubiquitinated peptides permit the study of the catalytic activities of deubiquitinases. We synthesized, peptides derived from ubiquitinated lysines at the surface of mdm2, and characterized them biophysically. We used a series of 15 deubiquitinase enzymes to test the use and the practicability of this approach. The method could be adapted to any peptide susceptible to ubiquitination. Variations in length and in sequence would permit the study of the Deubiquitinase/peptide recognition rules.