Chitinolytic β-N-acetyl-D-hexosaminidase is a branch of the GH20 (glycoside hydrolase family 20) β-N-acetyl-D-hexosaminidases that is only distributed in insects and micro-organisms, and is therefore a potential target for the action of insecticides. PUGNAc [O-(2-acetamido-2-deoxy-D-glucopyransylidene)-amino-N-phenylcarbamate] was initially identified as an inhibitor against GH20 β-N-acetyl-D-hexosaminidases. So far no crystal structure of PUGNAc in complex with any GH20 β-N-acetyl-D-hexosaminidase has been reported. We show in the present study that the sensitivities of chitinolytic β-N-acetyl-D-hexosaminidases towards PUGNAc can vary by 100-fold, with the order being OfHex1 (Ostrinia furnacalis β-N-acetyl-D-hexosaminidase)<SmCHB (Serratia marcescens chitobiase)<SpHex (Streptomyces plicatus β-N-acetyl-D-hexosaminidase). To explain this difference, the crystal structures of wild-type OfHex1 as well as mutant OfHex1(V327G) in complex with PUGNAc were determined at 2.0 Å (1 Å=0.1 nm) and 2.3 Å resolutions and aligned with the complex structures of SpHex and SmCHB. The results showed that the sensitivities of these enzymes to PUGNAc were determined by the active pocket size, with OfHex1 having the largest but narrowest entrance, whereas SpHex has the smallest entrance, suitable for holding the inhibitor, and SmCHB has the widest entrance. By widening the size of the active pocket entrance of OfHex1 through replacing the active site Val327 with a glycine residue, the sensitivity of OfHex1 to PUGNAc became similar to that of SmCHB. The structural differences among chitinolytic β-N-acetyl-D-hexosaminidases leading to different sensitivities to PUGNAc may be useful for developing species-specific pesticides and bactericides.
Skip Nav Destination
Article navigation
September 2011
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
August 26 2011
Active-pocket size differentiating insectile from bacterial chitinolytic β-N-acetyl-D-hexosaminidases
Tian Liu;
Tian Liu
1
*School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China
Search for other works by this author on:
Haitao Zhang;
Haitao Zhang
1
†State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China
Search for other works by this author on:
Fengyi Liu;
Fengyi Liu
*School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China
Search for other works by this author on:
Lei Chen;
Lei Chen
*School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China
Search for other works by this author on:
Xu Shen;
Xu Shen
2
†State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China
2Correspondence may be addressed to either of these authors (email xshen@mail.shcnc.ac.cn or qingyang@dlut.edu.cn).
Search for other works by this author on:
Qing Yang
Qing Yang
2
*School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China
2Correspondence may be addressed to either of these authors (email xshen@mail.shcnc.ac.cn or qingyang@dlut.edu.cn).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
March 01 2011
Revision Received:
June 08 2011
Accepted:
June 21 2011
Accepted Manuscript online:
June 21 2011
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2011 Biochemical Society
2011
Biochem J (2011) 438 (3): 467–474.
Article history
Received:
March 01 2011
Revision Received:
June 08 2011
Accepted:
June 21 2011
Accepted Manuscript online:
June 21 2011
Citation
Tian Liu, Haitao Zhang, Fengyi Liu, Lei Chen, Xu Shen, Qing Yang; Active-pocket size differentiating insectile from bacterial chitinolytic β-N-acetyl-D-hexosaminidases. Biochem J 15 September 2011; 438 (3): 467–474. doi: https://doi.org/10.1042/BJ20110390
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.