Abstract
Peritrophic membranes (PMs) are composed of chitin and protein. Chitin and protein play important roles in the structural formation and function of the PM. A new type I PM protein, HoCBP76, was identified from the Holotrichia oblita. HoCBP76 was shown as a 62.3 kDa protein by SDS-PAGE analysis and appeard to be associated with the PM throughout its entire length. In H. oblita larvae, the midgut is the only tissue where HoCBP76 could be detected during the feeding period of the larvae. The predicted amino acid sequence indicates that it contains seven tandem chitin binding domains belonging to the peritrophin-A family. HoCBP76 has chitin binding activity and is strongly associated with the PM. The HoCBP76 was not a mucin-like glycoprotein, and the consensus of conserved cysteines appeared to be CX13–17CX5CX9CX12CX7C. Western blot analysis showed that the abundance of HoCBP76 in the anterior, middle and posterior regions of the midgut was similar, indicating that HoCBP76 was secreted by the whole midgut epithelium, and confirmed the H. oblita PM belonged to the Type I PM. Immunolocalization analysis showed that HoCBP76 was mainly localized in the PM. The HoCBP76 is the first PM protein found in the H. oblita; however, its biochemical and physiological functions require further investigation.
Highlights
At one or more stages of the life cycle in most insects, peritrophic membranes (PMs) surround the food bolus in the digestive tract, facilitate food digestion and provide protection for midgut epithelial cells from invasion by viruses, bacteria and other pathogens
chitin-binding domains (CBDs) was first characterized in the peritrophic membrane protein peritrophin-44 from Lucilia cuprina, which was confirmed to bind to chitin within the PM [16]
Western blot analysis confirmed that secreted the H. oblita PM belonged to the Type Ι PM, which was similar to the observations made in other Coleoptera insects, the Diabrotica undecimpunctata (Coleoptera: Chrysomelidae), which PM was synthesized along the length of the midgut epithelium, and secreted into the interstices between the microvilli of the brush border [11]
Summary
Xiaomin Liu 1,3, Jie Li 4, Wei Guo 2,3,*, Ruijun Li 3, Dan Zhao 3 and Xinna Li 3. Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206, China. College of Plant Protection, Agricultural University of Hebei/Biological Control Centre of. Received: 29 January 2014; in revised form: 20 March 2014 / Accepted: 3 April 2014 /
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