New α-adrenergic property for synthetic MTβ and CM-3 three-finger fold toxins from black mamba

Toxicon. 2013 Dec 1:75:160-7. doi: 10.1016/j.toxicon.2013.04.017. Epub 2013 May 3.

Abstract

Despite their isolation more than fifteen years ago from the venom of the African mamba Dendroaspis polylepis, very few data are known on the functional activity of MTβ and CM-3 toxins. MTβ was initially classified as a muscarinic toxin interacting non-selectively and with low affinity with the five muscarinic receptor subtypes while no biological function was determined for CM-3. Recent results highlight the multifunctional activity of three-finger fold toxins for muscarinic and adrenergic receptors and reveal some discrepancies in the pharmacological profiles of their venom-purified and synthetic forms. Here, we report the pharmacological characterization of chemically-synthesized MTβ and CM-3 toxins on nine subtypes of muscarinic and adrenergic receptors and demonstrate their high potency for α-adrenoceptors and in particular a sub-nanomolar affinity for the α1A-subtype. Strikingly, no or very weak affinity were found for muscarinic receptors, highlighting that pharmacological characterizations of venom-purified peptides may be risky due to possible contaminations. The biological profile of these two homologous toxins looks like that one previously reported for the Dendroaspis angusticeps ρ-Da1a toxin. Nevertheless, MTβ and CM-3 interact more potently than ρ-Da1a with α1B- and α1D-AR subtypes. A computational analysis of the stability of the MTβ structure suggests that mutation S38I, could be involved in this gain in function.

Keywords: 2-(6-Chloro-1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate; Adrenergic toxin; Adrenoceptor; G-protein coupled receptor; GPCR; GPCRs; HCTU; MTs; Muscarinic receptor; Muscarinic toxin; N-methy-2-lpyrrolidone; N-methylmorpholine; N-methylscopolamine; NMM; NMP; NMS; Three-finger fold toxin; mAChRs; muscarinic acetylcholine receptors; muscarinic toxins.

MeSH terms

  • Adrenergic Agents / chemical synthesis
  • Adrenergic Agents / chemistry*
  • Adrenergic Agents / isolation & purification
  • Amino Acid Sequence
  • Animals
  • Circular Dichroism
  • Elapidae*
  • Molecular Sequence Data
  • Protein Conformation
  • Receptors, Adrenergic / metabolism
  • Receptors, Muscarinic / metabolism
  • Snake Venoms / chemical synthesis
  • Snake Venoms / chemistry*
  • Toxins, Biological / chemical synthesis
  • Toxins, Biological / chemistry*
  • Toxins, Biological / isolation & purification

Substances

  • Adrenergic Agents
  • Receptors, Adrenergic
  • Receptors, Muscarinic
  • Snake Venoms
  • Toxins, Biological