Scorpion Venom

Analysis

Powdered venom was prepared in suspension at a concentration of 1000 ppm. The value was determined with the Cond meter SevenDirect SD23 measuring device, and the of the scorpion liquid venom was determined as 5.67

HPLC Analysis

Powdered venoms were loaded into the HPLC (High Performance Liquid Chromatogra) instrument at a concentration of 1000 ppm. In Shimadzu Prominence-i LC2030 3D HPLC device, gradient program with a flow rate of 1mg/mL. The spectrum was taken to determine the maximum absorbance of the proteins in the venom. In these results, HPLC analysis was performed at 280 mm wavelength. Proteins in the venom were detected in fractions at different times (Figure 2). As a result of HPLC analysis, 18 different proteins/peptides were determined in the venom of Androctonus turkiyensis. The retention time of the fractions was indicated (Table 1).

Analysis of Scorpion Venom Peptides

The venom of the Androctonus turkiyensis was examined for possible peptides. When the existing peptides were analyzed, 21 polypeptides bound to potassium channels were detected.

This peptide, which is a-KTx 3.6 potassium channel inhibitor, inhibits the target channels indicated in Table 2. It disrupts the intralar sodium/potassium balance and drags to death.

Table 2. Sequences and possible target channels of peptides identified in Androctonus turkiyensis venom

Determination of Protein Concentration

The amount of protein contained in raw venom obtained from scorpions was determined by a colorimetric assay, Bicinchoninic acid (BCA). Samples were measured at 562 nm after half an hour of incubation at 37°C. Bovine serum albumin (BSA) was used as a standard at concentrations of 2000, 1500, 1000, 750, 500, 250, 125, 25 and 0 g/mL. (Figure 3) Samples and standard were measured in duplicate. By using the optical density (OD) values, the standard grawas created with the BSA results, the slope formula was calculated from Microsoft Excel software, and the grawas obtained with Graad Prism 8 software. Protein concentration of 5.3 $ 0.3 mg/mL protein concentration was determined for Androctonus turkiyensis, which was studied in duplicate.

SDS-PAGE Analysis

Electroretic separation of Androctonus turkiyensis venom was performed with a vertical electroresis system consisting of 4-12% acrylamide under reducing conditions. Lane 1: molecular weight standard (range 10-250 kDa), Lane 2: 5 ug Androctonus turkiyensis venom, Lane 3: 10 ug Androctonus turkiyensis venom, Lane 4: 15 ug Androctonus turkiyensis venom, Lane 5: 20 ug Androctonus turkiyensis venom. Marks indicated on the left are expressed in kDa. Lane 6: 30 ug Androctonus turkiyensis venom. Marks indicated on the left are expressed in kDa.After SDS-PAGE analysis, the presence of 5-75 kDa proteins was detected in Androctonus turkiyensis venom. However, it is seen that peptides are concentrated in 37 kDa, 25 kDa and 17 kDa weight of proteins in total scorpion venom, especially a protein/peptide density of 20 kDa.