Figure 9.

Analysis of the effect of different concentrations of SDS on the cleavage efficiency by five different enzymes, hPR-3, hNE, hCG, human thrombin and xPR-3. Consensus substrate sequences for the different enzymes were inserted in the 2x Trx substrates as shown in Figure 5. These substrates were then were used to analyze the effect of different SDS concentration on the cleavage efficiency by the five different enzymes. We used the substrate (VLLVSEVL) for hNE, the substrate (VLLFSEVL) for hCG, the substrate (VLLVSEVL) for hPR-3, the substrate (LTPRGVRL) for human thrombin and the substrate (VLLVSEVL) for Xenopus PR-3. Lane C for the different enzymes is a control without SDS. The concentrations of the SDS are then listed above the corresponding lane from 0.1 to 8%. The uncleared substrates have a molecular weight of ≈25 kDa and the cleaved substrates appear as two closely located bands with a size of 12–13 kDa. Due to the effect of the SDS to open the protein structure of the Trx molecules cleavage also occur within the Trx molecules, which results in additional bands of different molecular weights.