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5 July 2020 Identification and Characterization of Aminopeptidase-N as a Binding Protein for Cry3Aa in the Midgut of Monochamus alternatus (Coleoptera: Cerambycidae)
Yajie Guo, Rebeca Carballar-Lejarazú, Liangjing Sheng, Yan Fang, Shaozhen Wang, Guanghong Liang, Xia Hu, Rong Wang, Feiping Zhang, Songqing Wu
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Abstract

Bacillus thuringiensis Cry proteins have been widely used over the past decades for many different insect pests, which are safe for users and the environment. The coleopteran-specific Cry3Aa toxin from B. thuringiensis exhibits toxicity to the larvae of Monochamus alternatus. Receptors play a key role in the mechanisms underlying the toxic action of Cry. However, the binding receptor for Cry3Aa has yet to be identified in the midgut of M. alternatus larvae. Therefore, the aim of this study was to identify the receptor for Cry3Aa toxin in the brush border membrane vesicles (BBMVs) of M. alternatus larvae. Our results indicate that the Cry3Aa toxin binds to the BBMVs (Kd = 247 nM) of M. alternatus via a 107 kDa aminopeptidase N (APN) (Kd = 57 nM). In silico analysis of the APN protein predicted that an 18 amino acid sequence in the N-terminal acted as a signal peptide, and that the Asn residue, located at position 918 in the C-terminus is an anchored site for glycosyl phosphatidyl inositol. Further analysis showed that M. alternatus APN exhibits 75% homology to the APN from Anoplophora glabripenis. Our work, therefore, confirmed that APN, which is localized in the BBMVs in the midgut of M. alternatus larvae, acts as a binding protein for Cry3Aa toxins.

© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
Yajie Guo, Rebeca Carballar-Lejarazú, Liangjing Sheng, Yan Fang, Shaozhen Wang, Guanghong Liang, Xia Hu, Rong Wang, Feiping Zhang, and Songqing Wu "Identification and Characterization of Aminopeptidase-N as a Binding Protein for Cry3Aa in the Midgut of Monochamus alternatus (Coleoptera: Cerambycidae)," Journal of Economic Entomology 113(5), 2259-2268, (5 July 2020). https://doi.org/10.1093/jee/toaa130
Received: 9 January 2020; Accepted: 25 May 2020; Published: 5 July 2020
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KEYWORDS
aminopeptidase N
Cry3Aa toxin
Monochamus alternatus Hope
receptor
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