Meeting Abstract
Venomous marine cone snails (genus Conus) possess a high diversity of post-translationally modified peptide neurotoxins (conotoxins) . Our studies focus on the A-superfamily of conotoxin encoding genes from the fish-hunting cone snail, C. catus. A-superfamily members encode α-conotoxins and αA-conotoxins that cause flaccid paralysis by inhibiting nicotinic acetylcholine receptors (Azam and McIntosh 2009). In the pionoconus subgenus of fish-hunters that includes C. catus, the A-superfamily also encodes neuroexcitatory peptides (NEX) that induce rapid tetanic paralysis due to the overstimulation of neurons (Schulz 2004). NEX peptides have a novel target currently under investigation. Additionally, previous studies have shown that α4/7 conotoxins target a variety of neuronal-type nicotinic acetylcholine receptor (nAChR) subtypes, which could provide insight into the physiological roles of these receptor subtypes in pain, inflammation, nicotine addiction, Alzheimer’s disease, and Parkinson’s disease. Therefore, investigation of sequences encoding peptides in the A-superfamily from a diverse group of cone snail species would likely yield novel peptides for study. The focus of this research is on the isolation of gDNA from museum specimens for deep amplicon sequencing of A-superfamily members. Utilizing a comparative approach, we analyzed the sequences for novel NEX peptides and had select peptides synthesized. These peptides were folded into active conformations, and RP-HPLC was used to purify and quantify the samples. Protein folding was confirmed via mass spectrometry, and the activity of these peptides was investigated utilizing novel approaches developed to identify potential high affinity targets in the zebrafish model system.
