Comparative proteomic analysis of female and male venoms from the Mexican scorpion Centruroides limpidus: Novel components found

- Venom from male and female scorpion of Centruroides limpidus were fractionated separately.
- Proteomic analysis of the separated venoms components is reported.
- HPLC and two-dimensional gel separations shows the existence of differences.
- Two novel unknown components were identified in these venoms.
- The enzymatic activities of both types of venoms were assayed and demonstrated.

Venom from male and female scorpions of the species Centruroides limpidus were separated by HPLC and their molecular masses determined by mass spectrometry. The relative concentration of components eluting in equivalent retention times from the HPLC column shows some differences. A new peptide with 29 amino acids, cross-linked by three disulfide bonds was found in male scorpions and its structure determined. Another unknown peptide present in female venom, with sequence identity similar to K+-channel blocking peptide was isolated. This peptide contains 39 amino acid residues linked by three disulfide bonds. Due to sequence similarities, a systematic number (αKTx2.18) was assigned. Venom from male and female scorpions was separated by Sephadex G-50 gel filtration. Components of fraction I of this chromatogram were analyzed by two-dimensional gel electrophoresis and 41 spots were selected (20 from female and 21 from male). The spots were excised from the gel, enzymatically digested and sequenced by LC-MS/MS. This procedure allowed the identification of several proteins containing similar amino acid sequence of other known proteins registered on UniProt database. Among these proteins the presence of metalloproteinases (proteolytic enzymes), hyaluronidases and phosphatases were experimentally determined and shown to be present in both venom samples. The results shown here should help further work aimed at fully identification of the structure and function of venom components form C. limpidus male and female scorpions.