Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity

Background: Bacillus thuringiensis Cry toxins bind with different insect midgut proteins leading to toxin oligomerization, membrane insertion and pore formation. However, different Cry toxins had been shown to readily form high molecular weight oligomers or aggregates in solution in the absence of r...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Fang,Longfa, Wang,Bo, Zhou,Zishan, Yang,Sujuan, Shu,Changlong, Song,Fuping, Bravo,Alejandra, Soberón,Mario, Zhang,Jie
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2016
Materias:	Cry9Aa655 Disulfide bonds Oligomerization Insecticidal activity
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000300006
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:scielo:S0717-34582016000300006
record_format	dspace
spelling	oai:scielo:S0717-345820160003000062016-07-06Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activityFang,LongfaWang,BoZhou,ZishanYang,SujuanShu,ChanglongSong,FupingBravo,AlejandraSoberón,MarioZhang,Jie Cry9Aa655 Disulfide bonds Oligomerization Insecticidal activity Background: Bacillus thuringiensis Cry toxins bind with different insect midgut proteins leading to toxin oligomerization, membrane insertion and pore formation. However, different Cry toxins had been shown to readily form high molecular weight oligomers or aggregates in solution in the absence of receptor interaction. The role of Cry oligomers formed in solution remains uncertain. The Cry9A proteins show high toxicity against different Lepidoptera, and no-cross resistance with Cry1A. Results: Cry9Aa655 protein formed oligomers easily in solution mediated by disulfide bonds, according to SDS-PAGE analysis under non-reducing and reducing conditions. However, oligomerization is not observed if Cry9Aa655 is activated with trypsin, suggesting that cysteine residues, C14 and C16, located in the N-terminal end that is processed during activation participate in this oligomerization. To determine the role of these residues on oligomerization and in toxicity single and double alanine substitution were constructed. In contrast to single C14A and C16A mutants, the double C14A-C16A mutant did not form oligomers in solution. Toxicity assays against Plutella xylostella showed that the C14A-C16A mutant had a similar insecticidal activity as the Cry9Aa655 protein indicating the oligomers of Cry9Aa formed in solution in the absence of receptor binding are not related with toxicity. Conclusions: The aggregation of Cry9Aa655 polypeptides was mediated by disulfide bonds. Cry9Aa655 C14 and C16C are involved in oligomerization in solution. These aggregate forms are not related to the mode of action of Cry9Aa leading to toxicity.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.19 n.3 20162016-05-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000300006en10.1016/j.ejbt.2016.02.005
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Cry9Aa655 Disulfide bonds Oligomerization Insecticidal activity
spellingShingle	Cry9Aa655 Disulfide bonds Oligomerization Insecticidal activity Fang,Longfa Wang,Bo Zhou,Zishan Yang,Sujuan Shu,Changlong Song,Fuping Bravo,Alejandra Soberón,Mario Zhang,Jie Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity
description	Background: Bacillus thuringiensis Cry toxins bind with different insect midgut proteins leading to toxin oligomerization, membrane insertion and pore formation. However, different Cry toxins had been shown to readily form high molecular weight oligomers or aggregates in solution in the absence of receptor interaction. The role of Cry oligomers formed in solution remains uncertain. The Cry9A proteins show high toxicity against different Lepidoptera, and no-cross resistance with Cry1A. Results: Cry9Aa655 protein formed oligomers easily in solution mediated by disulfide bonds, according to SDS-PAGE analysis under non-reducing and reducing conditions. However, oligomerization is not observed if Cry9Aa655 is activated with trypsin, suggesting that cysteine residues, C14 and C16, located in the N-terminal end that is processed during activation participate in this oligomerization. To determine the role of these residues on oligomerization and in toxicity single and double alanine substitution were constructed. In contrast to single C14A and C16A mutants, the double C14A-C16A mutant did not form oligomers in solution. Toxicity assays against Plutella xylostella showed that the C14A-C16A mutant had a similar insecticidal activity as the Cry9Aa655 protein indicating the oligomers of Cry9Aa formed in solution in the absence of receptor binding are not related with toxicity. Conclusions: The aggregation of Cry9Aa655 polypeptides was mediated by disulfide bonds. Cry9Aa655 C14 and C16C are involved in oligomerization in solution. These aggregate forms are not related to the mode of action of Cry9Aa leading to toxicity.
author	Fang,Longfa Wang,Bo Zhou,Zishan Yang,Sujuan Shu,Changlong Song,Fuping Bravo,Alejandra Soberón,Mario Zhang,Jie
author_facet	Fang,Longfa Wang,Bo Zhou,Zishan Yang,Sujuan Shu,Changlong Song,Fuping Bravo,Alejandra Soberón,Mario Zhang,Jie
author_sort	Fang,Longfa
title	Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity
title_short	Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity
title_full	Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity
title_fullStr	Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity
title_full_unstemmed	Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity
title_sort	oligomerization of cry9aa in solution without receptor binding, is not related with insecticidal activity
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2016
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000300006
work_keys_str_mv	AT fanglongfa oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT wangbo oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT zhouzishan oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT yangsujuan oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT shuchanglong oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT songfuping oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT bravoalejandra oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT soberonmario oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity AT zhangjie oligomerizationofcry9aainsolutionwithoutreceptorbindingisnotrelatedwithinsecticidalactivity
_version_	1718441928149696512

Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity

Ejemplares similares