PXD055041-1

PXD055041 is an original dataset announced via ProteomeXchange.

Title	Cry1Ac Toxin Binding in the Velvetbean Caterpillar Anticarsia gemmatalis: Study of Midgut Aminopeptidases N
Description	The velvetbean caterpillar Anticarsia gemmatalis is one of the main soybean defoliators in Brazil. Currently, the main biopesticide used to control not only A. gemmatalis but also other insect pests worldwide is the bacteria Bacillus thuringiensis (Bt), which produces entomopathogenic Crystal toxins (Cry) that act in the midgut of susceptible insects, leading them to death. The mode of action of Cry toxins in the midgut involves binding to specific receptors present on the brush border of epithelial cells such as aminopeptidase N (APN), alkaline phosphatase (ALP), cadherin, and others. Mutations in these receptors, among other factors, may be involved in the development of resistance; identification of functional Cry receptors in the midgut of A. gemmatalis is crucial to develop effective strategies to overcome this possible scenario. This study’s goal is to characterize APNs of A. gemmatalis and identify a receptor for Cry1Ac in the midgut. The interaction of Bt spores with the midgut epithelium was observed in situ by immunohistochemistry and total aminopeptidase activity was estimated in brush border membrane vesicle (BBMV) samples, presenting higher activity in challenged individuals than in control ones. Ten APN sequences were found in a A. gemmatalis’ transcriptome and subjected to different in silico analysis, such as phylogenetic tree, multiple sequence alignment and identification of signal peptide, activity domains and GPI-anchor signal. BBMV proteins from 5th instar larvae were submitted to a ligand blotting using activated Cry toxin and a commercial anti-Cry polyclonal antibody; corresponding bands of proteins that showed binding to Cry toxin were excised from the SDS-PAGE gel and subjected to mass spectrometry analysis, which resulted in the identification of seven of those APNs. Quantitative PCR was realized to compare expression levels between individuals subjected to sublethal infection with Bt spores and control ones, presenting up- and downregulations upon Bt infection. From these results, we can infer that aminopeptidases N in A. gemmatalis could be involved in the mode of action of Cry toxins in its larval stage.
HostingRepository	PRIDE
AnnounceDate	2025-05-07
AnnouncementXML	Submission_2025-05-07_01:02:27.685.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Luis Felipe Ramos
SpeciesList	scientific name: Anticarsia gemmatalis; NCBI TaxID: 129554;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2024-08-22 06:04:36	ID requested
⏵ 1	2025-05-07 01:02:28	announced

Lanzaro MD, Padilha I, Ramos LFC, Mendez APG, Menezes A, Silva YM, Martins MR, Junqueira M, Nogueira FCS, AnoBom CD, Dias GM, Gomes FM, Oliveira DMP, : study of midgut aminopeptidases N. Front Physiol, 15():1484489(2024) [pubmed]

10.3389/fphys.2024.1484489;

submitter keyword: Anticarsia gemmatalis, Cry Toxin, Bacillus thuringiensis, Receptor,Aminopeptidase N

Danielle Oliveira
contact affiliation	Laboratório de Bioquímica Estrutural de Proteínas, Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro - Rio de Janeiro, Brasil
contact email	danioliveira@iq.ufrj.br
lab head
Luis Felipe Ramos
contact affiliation	Federal University of Rio de Janeiro
contact email	luisfelipecostaramos@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD055041
     1. Label: PRIDE project
     2. Name: Cry1Ac Toxin Binding in the Velvetbean Caterpillar Anticarsia gemmatalis: Study of Midgut Aminopeptidases N