Abstract
An aromatic amino acid, Tyr or Trp, located in the esterase active site wall, is highly conserved, with hyperthermophilic esterases showing preference for Tyr and lower temperature esterases showing preference for Trp. In this study, we investigated the role of Tyr(182) in the active site wall of hyperthermophilic esterase EstE1. Mutation of Tyr to Phe or Ala had a moderate effect on EstE1 thermal stability. However, a small-to-large mutation such as Tyr to His or Trp had a devastating effect on thermal stability. All mutant EstE1 enzymes showed reduced catalytic rates and enhanced substrate affinities as compared with wild-type EstE1. Hydrogen bond formation involving Tyr(182) was unimportant for maintaining EstE1 thermal stability, as the EstE1 structure is already adapted to high temperatures via increased intramolecular interactions. However, removal of hydrogen bond from Tyr(182) significantly decreased EstE1 catalytic activity, suggesting its role in stabilization of the active site. These results suggest that Tyr is preferred over a similarly sized Phe residue or bulky His or Trp residue in the active site walls of hyperthermophilic esterases for stabilizing the active site and regulating catalytic activity at high tempe...Continue Reading
References
Jul 7, 1999·Acta Crystallographica. Section D, Biological Crystallography·G De SimoneV Pavone
Feb 16, 2002·Journal of Molecular Biology·G De SimoneC Pedone
Nov 18, 2003·The Journal of Biological Chemistry·Giuseppina De SimoneGiuseppe Manco
Apr 22, 2004·The Journal of Biological Chemistry·Euiyoung Bae, George N Phillips
Feb 5, 2005·Applied and Environmental Microbiology·Jin-Kyu RheeJong-Won Oh
Apr 7, 2006·Applied and Environmental Microbiology·Jin-Kyu RheeJong-Won Oh
Jul 13, 2007·BMC Structural Biology·Jung-Sue ByunHyun-Soo Cho
Jun 11, 2008·Methods in Molecular Biology·Joanne E NettleshipArie Geerlof
Jun 1, 2011·Nucleic Acids Research·Aurélien GrosdidierOlivier Michielin
Jun 4, 2011·PloS One·Xiangdong ZhengHai Pang
Feb 9, 2012·Biotechnology Letters·Seunghee HongSei-Heon Jang
May 26, 2012·Journal of Bacteriology·Sei-Heon JangChangWoo Lee
Jul 4, 2012·The FEBS Journal·Clement AngkawidjajaShigenori Kanaya
Oct 31, 2012·International Journal of Molecular Sciences·Caroline Struvay, Georges Feller
Nov 3, 2012·Applied Biochemistry and Biotechnology·Changwoo LeeSei-Heon Jang
Feb 26, 2013·Bioresource Technology·Bao-Di MaJian-He Xu
Mar 29, 2014·Biochimica Et Biophysica Acta·Jerusha BoyineniSei-Heon Jang
Apr 1, 2014·Biochemical and Biophysical Research Communications·Shuai DouJian-He Xu
May 8, 2014·The Protein Journal·Md Abul Kashem TangKeiichi Watanabe
Jul 22, 2014·The Journal of Biological Chemistry·Kazuhiro OharaToru Nakayama
Dec 5, 2014·Extremophiles : Life Under Extreme Conditions·Jonathan MaiangwaThean Chor Leow
Feb 26, 2015·Biochemistry·Qi ChenJian-He Xu
Citations
Jun 5, 2016·Biochemistry·Ngoc TruongvanChangWoo Lee
Oct 13, 2018·Protein Science : a Publication of the Protein Society·Polytimi S DimitriouKonstantin Denessiouk
Feb 23, 2020·PloS One·Alexander DenesyukKonstantin Denessiouk
Jul 25, 2019·Extremophiles : Life Under Extreme Conditions·VinayKumar DachuriChangWoo Lee
Sep 24, 2020·Biochimica Et Biophysica Acta. Proteins and Proteomics·Kiet N TranChangWoo Lee
Dec 8, 2017·ACS Omega·Ayesha KashifChangWoo Lee