Apoptosis-inducing protein, AIP, from parasite-infected fish induces apoptosis in mammalian cells by two different molecular mechanisms

Abstract

AIP (apoptosis-inducing protein) is a protein purified and cloned from Chub mackerel infected with the larval nematode, Anisakis simplex, which induces apoptosis in various mammalian cells including human tumor cell lines. AIP has shown structural and functional homology to Lamino acid oxidase (LAO) which oxidizes several L-amino acids including L-lysine and AIP-induced apoptosis has been suggested to be mediated by H 2 O 2 generated by LAO activity of AIP. In this study, we confirmed that recombinant AIP generated enough H 2 O 2 in culture medium to induce rapid apoptosis in cells and this apoptosis was clearly inhibited by co-cultivation with antioxidants such as catalase and Nacetyl-cysteine. Surprisingly, however, we found that AIP still could induce H 2 O 2 -independent apoptosis more slowly than H 2 O 2 -dependent one in HL-60 cells even in the presence of antioxidants. In addition, the HL-60-derived cell line HP100-1, which is a H 2 O 2 -resistant variant, underwent apoptosis on treatment with AIP with a similar delayed time course. The latter apoptosis was completely blocked by addition of L-lysine to the culture medium, which is the best substrate of AIP as LAO, indicating that decreased concentration of L-lysine in the culture medium by AIPtreatment induced apoptosis. We also showed that the both apoptosis by AIP were associated with the release of cytochrome c from mitochondria and activation of caspase-9, and overexpressed Bcl-2 could inhibit both of the AIPinduced apoptosis. These results indicate that AIP induces apoptosis in cells by two distinct mechanisms; one rapid and mediated by H 2 O 2 , the other delayed and mediated by deprivation of L-lysine, both of which utilize caspase-9/ cytochrome c system. Cell Death and Differentiation (2001) 8, 298 ± 307.

Molecular profiling of the gilthead sea bream (Sparus aurata L.) response to chronic exposure to the myxosporean parasite Enteromyxum leei

“…The skin culture fluid from channel catfish resistant to the ciliate Ichthyophthirius multifiliis induces apoptosis of parasite theronts (Xu et al, 2005). Furthermore apoptosis-inducing protein, a protein purified from chub mackerel infected with the nematode Anisakis simplex, provokes apoptosis in various mammalian cell lines (Murakawa et al, 2001). These apoptotic adjustments may also reflect the host's attempt to reduce immunopathology associated with unwanted activity of effector leukocytes and return to a status of cell/tissue homeostasis.…”

Protease Activity of Procaspase-8 Is Essential for Cell Survival by Inhibiting Both Apoptotic and Nonapoptotic Cell Death Dependent on Receptor-interacting Protein Kinase 1 (RIP1) and RIP3

“…Results obtained by trypan blue exclusion method were essentially same as those by LDL release method. The DNA ladder assay was performed as described (33). To quantify apoptotic cells, cells were stained with propidium iodide (PI) and percentage of sub-diploid (sub-G1) cells was measured by flow cytometric analysis as previously described (30).…”

Induction of apoptosis in yeast by L‐amino acid oxidase from the Malayan pit viper Calloselasma rhodostoma

“…We have reported that the depletion of an essential amino acid can trigger apoptotic cell death in yeast (Eisler et al, 2004). Other members of the LAAO family, such as achacin from the body surface mucus of the giant African snail (Achatina fulica Ferussac) and the apoptosis-inducing protein (AIP) from parasiteinfected fish, deplete arginine, lysine, tryptophan and tyrosine, or only lysine, from mammalian cell culture medium (Kanzawa et al, 2004;Murakawa in the presence of catalase (1.5 µM) for 30 min. The cells were then washed three times with PBS and digested with proteinase K (0.2 mg/ml).…”