Multiple forms of alpha-2 macroglobulin in shrimp Fenneropenaeus chinesis and their transcriptional response to WSSV or Vibrio pathogen infection

Abstract

Alpha-2 macroglobulin (A2M) is a non-specific protease inhibitor involved in host defense. By full length cloning and sequencing we identified three distinct cDNAs for A2M in Chinese shrimp Fenneropenaeus chinesis, designated FcA2M-1, FcA2M-2 and FcA2M-3, respectively. Expression profiles in normal tissues as well as tissues after challenge by white spot syndrome virus (WSSV) and Vibrio pathogen were conducted for FcA2M-1 and FcA2M-2. The FcA2M-1 and FcA2M-2 cDNAs encode proteins with 1501 or 1502 amino acids, respectively, containing the typical conserved domain architecture of A2M. Similar to complement component C3, FcA2M-2 has a catalytic histidine, which may confer opsonic properties on this shrimp A2M. Six variants in the bait region were found in FcA2M-2 responding differently to Vibrio challenge, thereby widening the spectrum of inhibition and the diversity of immune recognition. FcA2M-1 and FcA2M-3, as well as most other protostomia invertebrate A2Ms identified so far, have a serine residue in the catalytic histidine position instead of the conserved asparagine residue found in vertebrate A2Ms. This, as inferred from a carp C3 molecule in which the catalytic histidine is substituted by a serine, suggests A2Ms in lower invertebrates possibly bear C3-like opsonic activity. These FcA2Ms showed much lower similarity to each other than to the A2Ms in other shrimp species, further supported by pylogenetic analysis. FcA2M-1 was found to be expressed most highly in hemocytes and lymphoid organ, while FcA2M-2 was expressed most highly in the heart and lymphoid organ, with the lowest expression in hemocytes. Challenge by WSSV or Vibrio pathogen increased the FcA2M-1 mRNA level in both hemocytes and lymphoid organ. After challenge, FcA2M-2 showed up-regulation in lymphoid organ but not in hemocytes. These expression features indicate that the different types of A2M in F. chinesis carry out different functions and that they are not simply functionally redundant.