Separation and identification of haemolymph proteins involved in immune response to Nosema pernyi infection in Antheraea pernyi (Lepidoptera: Saturniidae) larvae

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This study was aimed to preliminarily identify the proteins involved in the early immune response in the haemolymph from Antheraea pernyi larvae infected by Nosema pernyi. The 5th instar female larvae of A. pernyi were fed with N. pernyi spores and the proteins in the haemolymph were separated by SDS-PAGE, and the proteome profiles of the differential protein bands were further analyzed and identified by LC-MS/MS and proteomics analysis. One-dimensional SDS-PAGE analysis revealed that protein bands at 44 kD (AP44) and 28 kD (AP28) were enriched in the haemolymph of the female larvae at 144 h after feeding with the spores of N. pernyi. A total of 117 non-redundant proteins were identified by LC-MS/MS and searching the COG database, including 52 unique proteins from AP28, 53 unique proteins from AP44, and 12 proteins from both of them. Among them, 29 proteins are immune-related, including 15 proteins from AP28, such as heat shock proteins, ubiquitin-like proteins, ubiquitin-conjugating enzyme E2, juvenile hormone epoxide hydrolase, microtubule-associated protein RP/EB family member 3, lysozyme-like protein, ADP-ribosylation factor, putative defense protein and peptidoglycan recognition protein-like protein, and 10 proteins from AP44, such as DRK, prophenoloxidase and hemolin. Four proteins were identified from both AP28 and AP44, including heat shock protein hsp21.4, prophenoloxidase and basic attacin. The results provide the basis for further study on the mechanism of immune response and defense against N. pernyi infection in A. pernyi.

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