Critical amino acids for the insecticidal activity of Vip3Af from Bacillus thuringiensis: Inference on structural aspects

Scientific Reports
Núria BanyulsJuan Ferré

Abstract

Vip3 vegetative insecticidal proteins from Bacillus thuringiensis are an important tool for crop protection against caterpillar pests in IPM strategies. While there is wide consensus on their general mode of action, the details of their mode of action are not completely elucidated and their structure remains unknown. In this work the alanine scanning technique was performed on 558 out of the total of 788 amino acids of the Vip3Af1 protein. From the 558 residue substitutions, 19 impaired protein expression and other 19 substitutions severely compromised the insecticidal activity against Spodoptera frugiperda. The latter 19 substitutions mainly clustered in two regions of the protein sequence (amino acids 167-272 and amino acids 689-741). Most of these substitutions also decreased the activity to Agrotis segetum. The characterisation of the sensitivity to proteases of the mutant proteins displaying decreased insecticidal activity revealed 6 different band patterns as evaluated by SDS-PAGE. The study of the intrinsic fluorescence of most selected mutants revealed only slight shifts in the emission peak, likely indicating only minor changes in the tertiary structure. An in silico modelled 3D structure of Vip3Af1 is proposed for the...Continue Reading

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Citations

Aug 10, 2020·Nature Communications·Rafael Núñez-RamírezErnesto Arias-Palomo
May 16, 2021·Nature Communications·Matthew J ByrneRebecca F Thompson

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Datasets Mentioned

BETA
AJ872070.1
AAC37036
AAR40284
CAI43276
CAI43277
CAI43275
ACL97352
ABH10614
KC156693
KF826717

Methods Mentioned

BETA
affinity purification
gel filtration
Fluorescence
PCR
size exclusion chromatography
transgenic

Software Mentioned

SIB Compute pI / Mw
DiANNA
PyMOL
Conseq
Robetta
TotalLab
msa
POLO
PC
Geneious

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