title: New semi-synthesis of ubiquitin C-terminal conjugate with 7-amino-4-methylcoumarin.
abstract: The ligation of peptide hydrazides at a Gly site carrying a removal auxiliary was found to be an efficient process. This technology was successfully used for the synthesis of ubiquitin C-terminal conjugates. Recombinant Ub(1-75)-NHNH2 was prepared through the hydrozinolysis of the Ub(1-75)-intein fusion protein. It was ligated with a glycine derivative modified with an acid-sensitive thiol auxiliary. The final acid treatment produced the desired bioactive ubiquitin conjugates in practical quantities. Thus, the method described here extends the protocols of expressed protein ligation.
authors.affiliation: Key Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University, Shenzhen Graduate School, Shenzhen, 518055, China.|Tsinghua-Peking Center for Life Sciences, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
authors.forename: Yi-Tong
authors.initials: YT
authors.lastname: Li
authors.affiliation: Tsinghua-Peking Center for Life Sciences, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Liang
authors.affiliation: Tsinghua-Peking Center for Life Sciences, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
authors.forename: Jia-Bin
authors.initials: JB
authors.lastname: Li
authors.affiliation: Tsinghua-Peking Center for Life Sciences, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
authors.forename: Ge-Min
authors.initials: GM
authors.lastname: Fang
authors.affiliation: Key Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University, Shenzhen Graduate School, Shenzhen, 518055, China.|Tsinghua-Peking Center for Life Sciences, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Tsinghua-Peking Center for Life Sciences, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
mesh_terms: D060326:Chemistry Techniques, Synthetic / Q000379:methods*; D003374:Coumarins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D015394:Molecular Structure; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D025822:Ubiquitin C / Q000138:chemical synthesis* / Q000737:chemistry
chemical_list: D003374:Coumarins; D011994:Recombinant Proteins; D025822:Ubiquitin C; C028743:7-amino-4-methylcoumarin
keywords: deubiquitinating enzymes; expressed protein ligation; native chemical ligation; peptide hydrazides; ubiquitin