The enzyme Cypridina luciferase (CLase) enables Cypridina luciferin to emit light efficiently through an oxidation reaction. The catalytic mechanism on the substrate of CLase has been studied, but the details remain to be clarified. Here, we examined the luminescence of Cypridina luciferin in the presence of several proteins with drug-binding ability. Luminescence measurements showed that the mixture of human plasma alpha 1-acid glycoprotein (hAGP) and Cypridina luciferin produced light. The total value of the luminescence intensity over 60 s was over 12.6-fold higher than those in the presence of ovalbumin, human serum albumin, or bovine serum albumin. In the presence of heat-treated hAGP, the luminescence intensity of Cypridina luciferin was lower than in the presence of intact hAGP. Chlorpromazine, which binds to hAGP, showed an inhibitory effect on the luminescence of Cypridina luciferin, both in the presence of hAGP and a recombinant CLase. Furthermore, BlastP analysis showed that hAGP had partial amino acid sequence similarity to known CLases in the region including amino acid residues involved in the drug-binding ability of hAGP. These findings indicate enzymological similarity between hAGP and CLase and provide insights into both the enzymological understanding of CLase and development of a luminescence detection method for hAGP.

Injection of shrimp with non-specific double-stranded RNA (dsRNA) of diverse lengths, sequences, and base compositions is known to induce non-specific immunity and protect against lethal disease, although the mechanisms are unclear. Previous shrimp studies examined the effects of non-specific RNA on particular pathways, while their global effects have not been examined. To understand the global effects of non-specific RNA in shrimp, we injected kuruma shrimp (Marsupenaeus japonicus) with a dsRNA and a small interfering RNA (siRNA) that is not specific to any gene in the shrimp genome and then examined global gene expression at 24 and 48 h with a microarray. For the non-specific RNA, we chose double-stranded green fluorescent protein (dsGFP) and siGFP because they are commonly used as mock controls and their effects on shrimp have not yet been studied. Injection of PBS was used as a control. The microarray results showed that many genes were up-regulated and some were down-regulated by dsGFP. In addition, dsGFP injection increased survival following WSSV challenge. The changes in expression for several genes were confirmed by quantitative PCR. The up-regulated genes included genes for eight immune-related proteins: c-type lectin 2, hemocyte homeostasis-associated protein, viral responsive protein, fibrinogen-related protein 1, sid-1 like protein, argonaute 2, Dicer 2, and heat shock protein 90. These results show that injection of shrimp with non-specific dsRNA hinders viral accumulation and prevents significant mortalities.

Lucensosergia lucens is a luminous marine shrimp that has been suggested to use a coelenterazine-dependent luminescence system. However, the genetic information related to the luminescence system is lacking. Our RNA-Seq analysis of this shrimp did not show the existence of known or homologous coelenterazine-dependent luciferase genes. Subsequent biochemical analyses suggested that the shrimp possessed unknown proteinaceous components for coelenterazine luminescence.