Diversity of Immune Response Proteins: Nickel-Isolated Sp185/333 Proteins from the Purple Sea Urchin
Open AccessEffective protection against pathogens requires hosts to generate diverse immune receptors to detect the widest possible range of pathogens and to induce the production of a wide range of immune effector proteins. Sea urchins lack an adaptive immune system and may have unique mechanisms to diversify their anti-pathogen proteins. Strongylocentrotus purpuratus, the California purple sea urchin, has an estimate of ~50 members of the Sp185/333 gene family, which encode a highly diversified repertoire of immune response proteins that are produced following bacterial infection. Up to 260 different variants of the Sp185/333 proteins may be present in a single sea urchin, based on differences in size and charge, plus variations in presence and amount of each variant depending on the individual animal and type of immune challenge. A subset of native Sp185/333 proteins can be isolated by affinity to metal ions because of their multiple histidines, resulting in 1-4 bands of unique molecular weight on standard Western blots, which vary depending on the individual sea urchin. Two dimensional gel electrophoresis (2DE) of nickel-isolated native Sp185/333 (Ni-natSp185/333) protein samples, followed by Western blot analysis to detect Ni-natSp185/333 in the nickel-isolated samples. The blots show that the subset of proteins of a few molecular weights (MWs) observed by 1DE are composed of a complex of variants that differ in isoelectric point (pI). These Ni-natSp185/333 MW/pI variants differ among individual sea urchins. Furthermore, the repertoire of Ni-natSp185/333 protein variants varies in response to the same type of immune challenge for individual sea urchins. An increased understanding of Ni-natSp185/333 protein diversification will broaden our understanding of innate immune diversification capabilities in animals such as sea urchins that survive solely on innate immunity.
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