Localization and diversity of 185/333 proteins from the purple sea urchin – unexpected protein-size range and protein expression in a new coelomocyte type

Abstract

The current paradigm proposes that the innate immune systems of invertebrates are much more complex than previously thought. The highly diverse 185/333 gene family in the purple sea urchin encodes a family of closely related proteins of varying length and sequence composition. Subsets of small phagocytes and polygonal cells express 185/333 proteins with localization on the surface of the small phagocytes and within perinuclear vesicles in both cell types. In short-term cultures, coelomocytes form small aggregates that progress to syncytia that are thought to be equivalent to encapsulation in vivo. These aggregates were found to be enriched for 185/333-positive (185/333+) small phagocytes. In response to lipopolysaccharide challenge, coelomocytes transiently increased, including frequencies of both 185/333+ and 185/333-negative (185/333–) small phagocytes and 185/333+ polygonal cells. The 185/333 proteins were present in a broad array of sizes, most of which were larger than that predicted from the cDNAs. Recombinant 185/333 proteins expressed in bacteria and insect cells were also larger than expected, suggesting that the proteins dimerize and multimerize. The diversity of the 185/333 proteins, their expression in response to immune challenge, and their cellular localization suggests this protein family and the small phagocytes have an important immunological role in the sea urchin.