ABSTRACT
Proteins containing the late embryogenesis abundant (LEA) motif comprise a conserved family, postulated to act as cell protectors. However, their function and mechanisms of action remain unclear. Here we show that PRELI, a mammalian LEA-containing homolog of yeast Ups1p, can associate with dynamin-like GTPase Optic Atrophy-1 (OPA1) and contribute to the maintenance of mitochondrial morphology. Accordingly, PRELI can uphold mitochondrial membrane potential (ΔΨ(m)) and enhance respiratory chain (RC) function, shown by its capacity to induce complex-I/NADH dehydrogenase and ATP synthase expression, increase oxygen consumption and reduce reactive oxygen species (ROS) production. PRELI can also inhibit cell death induced by STS, TNF-α or UV irradiation. Moreover, in vitro and in vivo dominant-negative overexpression of mutant PRELI/LEA(-) (lacking the LEA motif) and transient in vitro PRELI-specific knockdown can render lymphocytes vulnerable to apoptosis, cause mouse embryo lethality and revert the resistance of lymphoma cells to induced death. Collectively, these data support the long-presumed notion of LEA protein-dependent mechanisms of cytoprotection and suggest that PRELI interacts with OPA1 to maintain mitochondria structures intact, sustain balanced ion(-)/proton(+) gradients, promote oxidative phosphorylation reactions, regulate pro- and antiapoptotic protein traffic and enable cell responses to induced death. These findings may help to understand how bioenergetics is mechanistically connected with cell survival cues.
Subject(s)
Mitochondrial Proteins/chemistry , Mitochondrial Proteins/metabolism , Proteins/chemistry , Proteins/metabolism , Amino Acid Motifs , Animals , Apoptosis , Caspases/metabolism , Cell Line, Tumor , Cell Respiration , Enzyme Activation , GTP Phosphohydrolases/metabolism , Gene Knockdown Techniques , Humans , Membrane Potential, Mitochondrial , Mice , Mitochondria/enzymology , Mitochondria/ultrastructure , Mitochondrial Membranes/metabolism , Mitochondrial Membranes/ultrastructure , Mitochondrial Proteins/ultrastructure , Protein Binding , Protein Transport , Proteins/ultrastructure , Sequence Deletion , Structure-Activity RelationshipABSTRACT
Many snake venoms have been shown to be complex mixtures of pharmacologically important molecules, some of which have potential therapeutic value in the treatment of clot-induced ischemia, cancer and other human disorders. The literature contains many references on how venom and/or venom components are being used in medicine. Within the United States, there are 44 subspecies of poisonous snakes. Despite this rather vast diversity, 90% of the venom-related biomedical research conducted on native snakes found in the United States has been done on a limited number of the more common species. Since the venoms from most of the native species are not available or characterized, their composition and potential usefulness in medicine and applied biomedical research has not been explored. The Natural Toxins Research Center (NTRC) at Texas A&M University-Kingsville has developed a serpentarium that presently houses a population of over 250 snakes composed of 11 species and 20 subspecies. These snakes are cataloged on the Internet database along with their geographical location data, proteolytic activities, high performance liquid chromatography (HPLC) and electrophoretic titration (ET) profiles. Many of these snake venoms have never been characterized and few locale-specific differences within a species have been examined. These venoms can be queried through an on-line search routine. The database will be a useful starting point for anyone interested in isolating fibrinolytic enzymes, specific toxins, hemorrhagins, or other pharmacologically active proteins from snake venoms.
Subject(s)
Databases, Factual , Internet , Snake Venoms , Animals , Chromatography, High Pressure Liquid , Humans , Mice , Mice, Inbred BALB C , Snake Venoms/chemistry , Snake Venoms/isolation & purification , Snake Venoms/toxicity , Snakes/classification , United StatesABSTRACT
Snake venoms contain direct-acting fibrinolytic metalloproteinases (MMP) that could have important applications in medicine. Fibrinolytic enzymes isolated from venom can induce in vitro clot lysis by directly acting on a fibrin clot. The most ideal fibrinolytic enzyme would have high affinity for clots, dissolve clots directly without causing hemorrhage, and would not be neutralized in vivo by endogenous metalloproteinase inhibitors. The purpose of this study was to compare DEAE/HPLC venom profiles from Viperid snakes and identify fractions that contain fibrinolytic activity with no hemorrhagic activity and are not neutralized by animal sera. The sera selected were from four (Virginia opossum, Gray woodrat, Mexican ground squirrel, and Hispid cottonrat) animals known to neutralize hemorrhagic activity in snake venoms. Nineteen fractions from the Viperid venoms had fibrinolytic activity. Agkistrodon venom fractions contained the highest specific fibrinolytic activities. A. piscivorus leucostoma fraction 4 contained a high specific fibrinolytic activity, no hemorrhagic activity, and the fibrinolytic activity was not neutralized by the proteinase inhibitors of the four animal sera. A. contortrix laticinctus fraction 1 also had a high specific fibrinolytic activity and no hemorrhagic activity. However, the fibrinolytic activity was neutralized by Didelphis virginiana (Virginia opossum) serum.
