title: Stable activity of a deubiquitylating enzyme (Usp2-cc) in the presence of high concentrations of urea and its application to purify aggregation-prone peptides.
abstract: Chemical synthesis of long or aggregation-prone peptide has been problematic. Its biological production has an advantage in that point, but it often forms inclusion body which creates difficulties in recovery of targets. As a deubiquitylating enzyme (Usp2-cc) was shown in this study to maintain its activity even in the presence of up to 4M urea, target peptide was purified by a single step of chromatography after overexpression as inclusion body, solubilization in urea and cleavage by the enzyme from the fusion protein consisting of GroES (used for high expression and easy to handle), ubiquitin (as a cleavage site) and target peptide. This system is a convenient tool for production of peptides that are difficult to be chemically synthesized and biologically purified.
authors.affiliation: Research Center for Proteineous Materials, Department of Bio-materials Engineering, Chosun University, Gwanju 501-759, Republic of Korea.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Shahnawaz
authors.forename: Arjun
authors.initials: A
authors.lastname: Thapa
authors.forename: Il-Seon
authors.initials: IS
authors.lastname: Park
mesh_terms: D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D018835:Chaperonin 10 / Q000235:genetics / Q000378:metabolism; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability / Q000187:drug effects; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011485:Protein Binding; D025801:Ubiquitin / Q000378:metabolism*; D014508:Urea / Q000494:pharmacology*
chemical_list: D016229:Amyloid beta-Peptides; D018835:Chaperonin 10; D010455:Peptides; D025801:Ubiquitin; D014508:Urea; D010450:Endopeptidases