Abstract
Chitinase B from Serratia marcescens 2170 is one of the processive chitinases, and it has a linear path of aromatic amino acid residues on the surface and in the catalytic cleft. There are four surface-exposed residues lined-up towards the cleft, Y481, W479, W252, and Y240. The substitution of these residues with alanine causes a decrease in both the extent of the substrate binding and the hydrolytic activity (Katouno et al., 2004). Here, we examine the three mutants without losing the substrate-binding ability, Y240W, Y481W, and Y240W/Y481W. These mutants were prepared for a detailed analysis of the functions of Y240 and Y481, which showed a lower contribution to substrate binding than W479 and W252. The parameters for the binding of the three mutants to crystalline β-chitin were similar to those for the wild type. The hydrolytic activity of Y240W and Y240W/Y481W against crystalline β-chitin was significantly decreased. However, the hydrolytic activity of Y481W was similar to that of the wild type, indicating some differences in the roles of Y240 and Y481 during the processive degradation of crystalline β-chitin. Taken together with the previous results, it was suggested that while Y240 and Y481 were required for the substrate...Continue Reading
References
May 18, 1981·Carbohydrate Research·H Yamada, T Imoto
Nov 1, 1995·Protein Science : a Publication of the Protein Society·C N PaceT Gray
Nov 26, 1997·Journal of Bacteriology·T WatanabeM Regue
Mar 21, 1998·Bioscience, Biotechnology, and Biochemistry·K SuzukiT Watanabe
May 24, 2000·Proceedings of the National Academy of Sciences of the United States of America·D M van AaltenR K Wierenga
Aug 2, 2001·Proceedings of the National Academy of Sciences of the United States of America·D M van AaltenV G Eijsink
Aug 28, 2001·The Journal of Biological Chemistry·T UchiyamaT Watanabe
Jan 30, 2003·Acta Crystallographica. Section D, Biological Crystallography·Yannis PapanikolauKyriacos Petratos
Aug 22, 2003·The Biochemical Journal·Takeshi WatanabeJunji Sugiyama
Aug 23, 2003·The Biochemical Journal·Nathan N AronsonPatrick Van Roey
Jul 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·Gennady ZolotnitskyYuval Shoham
Oct 22, 2004·Journal of Biochemistry·Fuminori KatounoTakeshi Watanabe
Feb 19, 2005·The Biochemical Journal·Eva-Lena HultJunji Sugiyama
Nov 23, 2006·Proceedings of the National Academy of Sciences of the United States of America·Svein J HornVincent G H Eijsink
Feb 27, 2009·The Journal of Biological Chemistry·Henrik ZakariassenVincent G H Eijsink
Oct 4, 2011·The Journal of Biological Chemistry·Christina M PayneGregg T Beckham
Feb 13, 2013·The FEBS Journal·Gustav Vaaje-KolstadVincent G H Eijsink
Feb 28, 2013·Biophysical Journal·Pavan K GhattyvenkatakrishnaXiaolin Cheng
Mar 28, 2013·The Journal of Biological Chemistry·Akihiko NakamuraMasahiro Samejima
Nov 26, 2013·Nucleic Acids Research·Vincent LombardBernard Henrissat
May 28, 2014·Current Opinion in Biotechnology·Gregg T BeckhamChristina M Payne
Jun 5, 2014·Nature Communications·Kiyohiko IgarashiMasahiro Samejima