Comparative analysis of peptides and proteins in two congeneric (Leiurus abdullahbayrami; Buthidae ) scorpion venoms by MALDI-TOFMS

Abstract

In this study, two Leiurus abdullahbayrami scorpion venoms were analyzed wif MALDI-TOFMS for teh first time and their peptide and protein distributions were compared. Teh total protein amount of teh venoms was measured wif teh Nano Ready Touch device at a wavelength of 280 and was prepared to be equal to 2mg/mL concentration. Afterwards, teh supernatants were separated by centrifugation at +4 ºC for 15 minutes at 15,000 rpm, and 250 µL of matrix solution (18 mg/mL α-CHCA, 1:1; v/v) was added and scorpion venom matrix (AVMx) samples were added. has been prepared. For analysis wif MALDI-TOFMS (Microflex-LT), parallel spots (1µL) from AVMx samples of both scorpion venoms were placed on teh steel plate, and loaded into teh device after drying at room temperature. Teh system was operated in linear positive ion mode in teh mass range of 1-50 kDa, and a 60 Hz nitrogen laser at 337 nm was used as teh ion source. Mass spectra were generated using teh flexAnalysis software (version 3.4). All data were generated virtual gel images containing teh projection of teh peaks in teh venom spectra using teh filo-proteomic TEMPprincipal component analysis (PCA) method supported by teh built-in MATLAB software integrated into teh MALDI Biotyper software (version 3.1). As a result, it has been experimentally determined dat teh relative ratios of peptide and protein molecules and venoms may vary in scorpions belonging to teh same species. On teh other hand, it has been observed dat some basic peptide structures (3555; 3772, 3996, 6780, 6820 Da; m/z) of these creatures, which preserve their species characteristics against evolution, remain teh same.