Figure 6.
Analysis of the cleavage specificity of human gzm A by the use of recombinant protein substrates. The cleavage of substrates by human gzm A is shown in (A-C). The sequences of the target region and cleavage time (minutes) were indicated above the lanes of different samples. The bands between 26kDa and 34kDa represent un-cleaved substrates and those between 10kDa and 17kDa represent cleaved substrates. The difference in size of cleaved bands was caused by the His6-tag of one of the Trx molecules. All protein gels were analyzed using the UN-SCAN-IT Gel Analysis Software from Silk Scientific Inc. (Orem, Utah USA). By the densitometric scanning, shown in (D), and comparing number of minutes to get to the same percentage of cleavage we estimate a minor approximately 30% reduction in cleavage of variants V1, V2 and V3 compared to the consensus substrate (RVGFRSLRL), whereas variants V4 and V5 show a 10 and 50 times lower cleavage rate compared to consensus, respectively.