Engineering of protease variants exhibiting altered substrate specificity

Biochemical and Biophysical Research Communications
Saravanan SellamuthuWoo Jin Park

Abstract

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.

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Citations

Apr 17, 2013·Proceedings of the National Academy of Sciences of the United States of America·Li YiBrent L Iverson
Jan 16, 2013·FEBS Letters·Qing LiBrent L Iverson
Jan 6, 2016·ACS Chemical Biology·Jennifer L GuerreroPatrick S Daugherty
Aug 28, 2009·Current Opinion in Biotechnology·Mark PogsonBrent L Iverson
Aug 9, 2016·Biotechnology and Bioengineering·Jennifer L GuerreroMichelle A O'Malley
Feb 16, 2012·Chembiochem : a European Journal of Chemical Biology·Tae Hyeon YooGeorge Georgiou
Jul 14, 2018·Bulletin of Experimental Biology and Medicine·T V BobikI V Smirnov

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