The crystal structure of the chitinase ChiA74 of Bacillus thuringiensis has a multidomain assembly

Scientific Reports
Estefania O Juárez-HernándezJ Eleazar Barboza-Corona

Abstract

There is no structural information about any chitinase synthesized by Bacillus thuringiensis, the most successful microbial insect larvicide used worldwide. In this study, we solved the 3D structure of the chitinase ChiA74 at 2.26 Å. The crystal structure shows that ChiA74 is composed of a modular arrangement formed by (i) a catalytic region (CD), (ii) a chitinase insertion domain (CID), (iii) a fibronectin type III domain (FnIII), and (iv) a chitin binding domain (CBD). The location of the CBD with respect to the CD has no structural similarity to other chitinases with known structures. The activity of a ChiA74 lacking its secretion signal peptide (ChiA74Δsp) and a truncated version lacking its CBD/FnIII domains (ChiA74Δsp-50) did not have statistical differences in activity against colloidal chitin. However, ChiA74Δsp exhibits 4.5 and 2.0 higher activity than versions lacking the CBD (ChiA74Δsp-60) and CBD/FnIII domains (ChiA74Δsp-50), respectively, when crystalline chitin was used as substrate. Our data suggest that the CBD might plays a significant role in crystalline chitin hydrolysis. We also demonstrated the importance of the catalytic E211 in the CD, as mutants ChiA74ΔspE211N and ChiA74ΔspD207N, E211N were inactive agai...Continue Reading

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Citations

Sep 8, 2019·World Journal of Microbiology & Biotechnology·Bao Le, Seung Hwan Yang
Apr 3, 2020·FEMS Microbiology Letters·Daisuke NikiJiro Arima
Jan 30, 2020·Frontiers in Microbiology·Sheila A Martínez-ZavalaJosé E Barboza-Corona
Apr 25, 2020·International Journal of Molecular Sciences·Piotr H MaleckiConstantinos E Vorgias
May 21, 2021·Frontiers in Molecular Biosciences·Yangchun Frank Chen, Yu Xia

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Methods Mentioned

BETA
electrophoresis

Software Mentioned

CCP4
AIMLESS
wwPDB Validation Server
UCSF Chimera
DIALS
Phaser
MatchMaker
Molprobity
ENDScript
LALING

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