PMID: 8962052Dec 10, 1996Paper

Protein engineering of Bacillus thuringiensis delta-endotoxin: mutations at domain II of CryIAb enhance receptor affinity and toxicity toward gypsy moth larvae

Proceedings of the National Academy of Sciences of the United States of America
F RajamohanD H Dean

Abstract

Substitutions or deletions of domain II loop residues of Bacillus thuringiensis delta-endotoxin CryIAb were constructed using site-directed mutagenesis techniques to investigate their functional roles in receptor binding and toxicity toward gypsy moth (Lymantria dispar). Substitution of loop 2 residue N372 with Ala or Gly (N372A, N372G) increased the toxicity against gypsy moth larvae 8-fold and enhanced binding affinity to gypsy moth midgut brush border membrane vesicles (BBMV) approximately 4-fold. Deletion of N372 (D3), however, substantially reduced toxicity (> 21 times) as well as binding affinity, suggesting that residue N372 is involved in receptor binding. Interestingly, a triple mutant, DF-1 (N372A, A282G and L283S), has a 36-fold increase in toxicity to gypsy moth neonates compared with wild-type toxin. The enhanced activity of DF-1 was correlated with higher binding affinity (18-fold) and binding site concentrations. Dissociation binding assays suggested that the off-rate of the BBMV-bound mutant toxins was similar to that of the wild type. However, DF-1 toxin bound 4 times more than the wild-type and N372A toxins, and it was directly correlated with binding affinity and potency. Protein blots of gypsy moth BBMV prob...Continue Reading

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Citations

Mar 5, 2008·Plant Physiology·John A Gatehouse
Jan 5, 2002·Applied and Environmental Microbiology·Periasamy BalasubramanianVaithilingam Sekar
Mar 28, 2002·Applied and Environmental Microbiology·Androulla GillilandDavid J Ellar
Sep 6, 2003·Applied and Environmental Microbiology·Mohd Amir F AbdullahDonald H Dean
Jul 8, 2005·Applied and Environmental Microbiology·Shogo AtsumiRyoichi Sato
Dec 13, 2005·Proceedings of the National Academy of Sciences of the United States of America·Claudia PérezAlejandra Bravo
Oct 24, 2014·Toxins·Benjamin R DeistBryony C Bonning
Feb 16, 2016·Journal of Basic Microbiology·Sameh SellamiKaïs Jamoussi
Jul 19, 2016·Nature Communications·Anilkumar GowdaJames A Baum
Mar 8, 2011·Plant Biotechnology Journal·Georgina SanahujaPaul Christou
Apr 3, 2012·Microbial Biotechnology·Alejandra BravoMario Soberón
Mar 4, 2000·Biotechnology and Bioengineering·B L Liu, Y M Tzeng
Jan 27, 2006·Protein Engineering, Design & Selection : PEDS·Xinyan Sylvia Liu, Donald H Dean
Mar 23, 2018·Scientific Reports·Mary-Carmen Torres-QuinteroAlejandra Bravo
Sep 22, 2018·Parasites & Vectors·Somphob LeetachewaSaengduen Moonsom
Nov 4, 2017·Scientific Reports·Tania Domínguez-FloresSusana Vílchez
Jul 2, 1999·Applied and Environmental Microbiology·C RangR Laprade

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