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    STUDIA PHYSICA - Issue no. 2 / 2004  
         
  Article:   STRUCTURAL ANALYSIS OF NANOMOLAR QUANTITIES OF NEUROPEPTIDES USING 2D-NMR.

Authors:  ELENA MATEI, HERMINSUL CANO, FRED PFLUEGER, FRANK MARI.
 
       
         
  Abstract:  Cone snails (genus Conus) are venomous and predatory marine mollusks whose venoms contain complex mixtures of neuroactive (conopeptides). Conopeptides have been recognized as versatile and powerful neuropharmacolo-gical agents. There are potentially ~ 100,000 different conopeptides that can target neuronal receptors, present in the venom of over 1000 species of cone snails. Every conopeptide has a highly specific target; binding of the conopeptide to its target leads to a biologically relevant change in neurophysiological function. After a laborious separation procedure, nanomole quantities of native conopeptides are usually obtained. For this study, we have selected two samples: i) 30 nmoles of -Conotoxin GI (ECCNPACGRHYSC; 2ľ7, 3ľ13), isolated from the venom of C. geographus, ii) 23 nanomoles of vexB, a 28-residue peptide with three disufide bonds isolated from Conus vexilum. Conformational analysis of nanomoles amount in aqueous solution of -conotoxin GI, a potent peptide antagonist of acetylcholine receptor (nAChR) and vexB, a potent blocker of Ca 2+ - ion channel, have been performed by two-dimensional NMR methods. Structural information of these conotoxins, was obtained from proton-proton internuclear distance constraints in combination with dynamical simulated annealing protocols. The limits of detection and other analytical parameters associated with the use of the nano-MAS and 1.7mm NMR probes in conopeptide research are evaluated.  
         
     
         
         
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