Abstract
Chitin-binding proteins (CBPs) are present in many species and they act in a variety of biological processes. We analyzed a Pteromalus puparum venom apparatus proteome and transcriptome and identified a partial gene encoding a possible CBP. Here, we report cloning a full-length cDNA of a sequence encoding a chitin-binding-like protein (PpCBP) from P. puparum, a pupal endoparasitoid of Pieris rapae. The cDNA encoded a 96-amino-acid protein, including a secretory signal peptide and a chitin-binding peritrophin-A domain. Phylogenetic analysis of chitin binding domains (CBDs) of cuticle proteins and peritrophic matrix proteins in selected insects revealed that the CBD of PpCBP clustered with the CBD of Nasonia vitripennis. The PpCBP is specifically expressed in the venom apparatus of P. puparum, mostly in the venom gland. PpCBP expression was highest at day one after adult eclosion and much lower for the following five days. We produced a recombinant PpCBP and binding assays showed the recombinant protein selectively binds chitin but not cellulose in vitro. We infer that PpCBP serves a structural role in the venom reservoir, or may be injected into the host to help wound healing of the host exoskeleton.
Highlights
Chitin-binding proteins (CBPs) are ubiquitous in organisms from viruses to invertebrates.Depending on the species, CBPs are involved in various biological functions, such as binding to chitin, antimicrobial activities, enhancing chitinolytic activity and hydrophobic surface sensing [1,2,3].In insects, CBPs are major components of insect cuticles and peritrophic membranes (PMs)
To establish the evolutionary relationships between PpCBP and other insect CBPs, a phylogenetic tree was constructed based on the amino acid sequences of the chitin binding domains (CBDs) (Figure 2)
Phylogenetic analysis of 24 CBDs from five insect species revealed the PpCBP CBD clustered with the CBD of NvCBP
Summary
Protein from Pupal Endoparasitoid Wasps, Pteromalus Puparum, Selectively Binds Chitin. Yu Zhu 1 , Xin-Hai Ye 2 , Yang Liu 1 , Zhi-Chao Yan 1 , David Stanley 3 , Gong-Yin Ye 1 and Qi Fang 1, *. Received: 13 October 2015; Accepted: 17 November 2015; Published: 30 November 2015
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