Neuronal calcium channel inhibitors: synthesis of ω-conotoxin GVIA and effects on 45Ca-uptake by synaptosomes

Journal ArticleWe previously described a 27-amino acid peptide neurotoxin from the venom of Conus geographus, wconotoxin GVIA, which inhibits neuronal voltage-activated calcium channels. In this paper we describe the total synthesis of ω-conotoxin GVIA and demonstrate that it efficiently blocks voltage-activated uptake of 46Ca by standard synaptosomal preparations from chick brain. Dihydropyridines do not block 46Ca uptake under these conditions

Conotoxins

Journal ArticleMany successful animal and plant families have developed distinctive biochemical strategies; one of the more unusual examples is found in a group of marine gastropods, the cone snails (Conus) (1). These animals have evolved a specialized biochemistry of small constrained peptides, the conotoxins. These peptides are the direct translation products of genes (2). However, because they are small enough for direct chemical synthesis and sufficiently constrained for three-dimensional conformation determination, conotoxins bridge protein chemistry and molecular genetics. Furthermore, the strategy that the cone snails have evolved over millions of years for the generation and design of an enormous array of small peptide ligands, each with high affinity and specificity for a particular receptor protein target, may be adaptable for use in vitro

Conotoxin MI: disulfide bonding and conformational states

Journal ArticleThe toxic peptide from Conus magus venom (conotoxin MI) is a 14-amino acid peptide (McIntosh, M., Cruz, L. J., Hunkapiller, M. W., Gray, W. R., and Olivera, B. M. (1982) Arch. Biochem. Biophys. 218, 329-334) which inhibits the acetylcholine ceptor. In this work we have confirmed the primary structure and established the disulfide bonding configuration (Cys 3-Cys 8; Cys 4-Cys 14) by direct chemical synthesis of the toxin with specific disulfide bridges

Protective effects of urocortin 2 against caerulein-induced acute pancreatitis.

Because little is known about the role of corticotropin-releasing factor (CRF) agonists in regulating responses in pancreatitis, we evaluated the effects of urocortin 2 (UCN2) and stressin1 in caerulein-induced acute pancreatitis (AP) model in rats. Male rats were pretreated with UCN2 or stressin1 for 30 min followed by induction of AP with supraphysiologic doses of caerulein. Serum amylase and lipase activity, pancreatic tissue necrosis, immune cell infiltrate, nuclear factor (NF)-κB activity, trypsin levels, and intracellular Ca2+ ([Ca2+]i) were ascertained. UCN2, but not stressin1 attenuated the severity of AP in rats. UCN2, but not stressin1, reduced serum amylase and lipase activity, cell necrosis and inflammatory cell infiltration in AP. NF-κB activity in pancreatic nuclear extracts increased in AP and UCN2 treatment reduced caerulein-induced increases in NF-κB activity by 42%. UCN2 treatment prevented caerulein-induced degradation of IκB-α in the cytosolic fraction as well as increased levels of p65 subunit of NF-κB in the cytosolic fraction. Pancreatic UCN2 levels decreased in AP compared with saline. UCN2 evoked [Ca2+]i responses in primary acinar cells and abolished caerulein-evoked [Ca2+]i responses at 0.1nM, and decreased by ~50% at 1.0nM caerulein. UCN2 stimulation resulted in redistribution of a portion of F-actin from the apical to the basolateral pole. UCN2 prevented the massive redistribution of F-actin observed with supraphysiologic doses of caerulein. UCN2, but not stressin1 attenuated severity of an experimental pancreatitis model. The protective effects of UCN2, including anti-inflammatory and anti-necrotic effects involve activation of the CRF2 receptor, [Ca2+]i signaling, and inhibition of NF-κB activity