Engineering of protease variants exhibiting altered substrate specificity

Autor:	Saravanan Sellamuthu, Bae Hyun Shin, Eui Seung Lee, Seong-Hwan Rho, Wangtaek Hwang, Yong Jae Lee, Hye-Eun Han, Jae Il Kim, Woo Jin Park
Rok vydání:	2008
Předmět:	Binding Sites Protein Conformation Biophysics 3C Viral Proteases Cell Biology Protein Engineering Biochemistry Substrate Specificity Cysteine Endopeptidases Viral Proteins Amino Acid Substitution Yeasts Directed Molecular Evolution Molecular Biology
Zdroj:	Biochemical and biophysical research communications. 371(1)
ISSN:	1090-2104
Popis:	By using an improved genetic screening system, variants of the HAV 3CP protease which exhibit altered P2 specificity were obtained. We randomly mutated the His145, Lys146, Lys147, and Leu155 residues that constitute the S2 pocket of 3CP and then isolated variants that preferred substrates with Gln over the original Thr at the P2 position using a yeast-based screening method. One of the isolated variants cleaved the Gln-containing peptide substrate more efficiently in vitro, proving the efficiency of our method in isolating engineered proteases with desired substrate selectivity.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d9a7e7bfe69472976957094b93c96424 https://pubmed.ncbi.nlm.nih.gov/18413229 Zobrazit plný text záznamu Full Text from ScienceDirect