Purification and Characterization of an Anti-Apoptotic Protein Isolated from Lonomia obliqua Hemolymph

Abstract

Previously it was reported that supplementation of insect cell culture with Lonomia obliqua hemolymph could extend culture longevity (Maranga et al. Biotechnol. Prog. 2003, 19, 58-63). In this work the anti-apoptotic properties of this hemolymph in Spodoptera frugiperda (Sf-9) cell culture were investigated. The presence or absence of apoptotic cells was characterized by light microscopy, flow cytometry, and agarose gel electrophoresis. Hemolymph was fractionated by several ion exchange and gel filtration chromatographic steps for identification of the compounds responsible for this effect. Fractions exhibiting a potent anti-apoptotic effect were isolated and tested in cell culture. A protein of about 51 kDa was identified, isolated, and tested for apoptosis inhibition. Addition of this purified protein to Sf-9 cultures was able to prevent apoptosis induced by nutrient depletion as well as by potent apoptosis chemical inducers such as Actinomycin D. This work confirms that the enhanced culture longevity obtained by supplementation with L. obliqua hemolymph is due to the presence of potent anti-apoptotic factors.

Toxins from Lonomia obliqua — Recombinant Production and Molecular Approach

“…This effect was attributed to the presence of three factors with different activities: a potential antiapoptotic factor, a growthpromoting factor, and an enzyme that hydrolyzes sucrose. Furthermore, an antiapoptotic protein of 51 kDa was purified from L. obliqua hemolymph [36]. This protein was able to prevent apoptosis in Sf-9 cell culture induced by nutrient deprivation and by Actinomycin D. Later reports [37,38] described in the hemolymph a potent antiviral activity against human virus.…”

“…The α-helices and β-sheets observed in Hyalophora cecropia hemolin (Protein Data Bank code 1BIH) and Losac are shaded in light and dark gray, respectively. Previously it was predicted that hemolin contain conserved regions and motifs (28,36). Conserved motifs are shown inside boxes according to the domain they belong.…”

The Medicinal Value of Biodiversity: New Hits to Fight Cancer

“…A more recent epigenetic role for parthenolide in cancer has been shown, by specifically inhibiting DNA methyltransferase 1 (DNMT1) and histone deacetylase 1 (HDAC1) activities, leading to DNA hypomethylation in vitro and in vivo (Liu et al, 2009;Ghantous et al, 2010). Also, the ability of plant secondary metabolites, such as vitamins, isothiocyanates, quercetin, catechins, resveratrol, epigallocatechin-3-gallate (EGCG), genistein and curcumin, to prevent and/or inhibit tumor growth, either alone or in combination with other anticancer drugs and/or other phytochemicals has been documented (de Souza et al, 2005;de Souza et al, 2006;Hemalswarya & Doble, 2006;de Souza Queiroz et al, 2007;Newman & Cragg, 2007;de Fátima et al, 2008a;Bailly, 2009;Camargo et al, 2011). In this respect, all-trans-retinoic acid (vitamin A) is now widely used for the treatment of myeloid leukemia (Sanz & Lo-Coco, 2011) and vitamin D is in clinical trial for this disease (Trump et al, 2006).…”

Improved replication of the baculovirus Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) in vitro using proteins from Lonomia obliqua hemolymph

“…Some studies have demonstrated the presence of pharmacologically active substances in hemolymphs from insects (Shiotsuki et al 2000;Yamamoto et al 1999;Guerrero et al 1990;Jiang et al 1999;Rosenfeld and Vanderberg 1998;Hamdaoui et al 1998;Maranga et al 2003;Lin et al 1998;Zhu et al 2000;Souza et al 2005). The supplementation of cell cultures with hemolymph proteins had a positive effect on viral replication (Rhee and Park 2000), and recombinant protein production (Woo et al 1997).…”