NM23 Is a CP-Binding Protein Involved in Infectious Hypodermal and Hematopoietic Necrosis Virus Infection in Shrimp

Abstract

The aim of this study was to identify the putative host cell receptor for Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) CP in the gill membrane of L. vannamei. Putative CP binding partners were screened first using a 2-dimensional Virus Overlay Protein Blot Assay (VOPBA) to probe isolated gill membrane proteins using recombinant CP. Putative binding partners were identified using mass spectrometry. A Phage Display Random Dodecapeptide Library was used to screen for dodecapeptides and motifs that bound to CP. Finally, putative binding pairs were confirmed using GST(glutathione-S-transferase) pulldown assays. 2-Dimensional VOPBA identified NM23 as a putative binding partner for IHHNV CP. GST pulldown experiments confirmed the direct interaction of NM23 and IHHNV CP. The phage display library was used to identify six groups of dodecapeptides that bound to CP. From these peptides, three characteristic binding motifs were identified, SW*Y, SKWV, and PQR. Interestingly, the SW*Y motif was also found in NM23. We are the first to implicate NM23 in IHHNV infection and postulate that it may bind to the CP using the SW*Y motif, although this remains to be confirmed.

Keywords

• IHHNV
• capsid protein
• gill membrane proteins
• receptors
• VOPBA
• NM23