Figure 4

SDS-PAGE and zymographic analyses of germinated sesame proteins elute from affinity columns. (A) The proteins extracted from the germinated sesame were collected after the purification with affinity columns, followed by SDS-PAGE analyses. Lane A, B and C were the pull-down proteins/extracted proteins via SiCYS, SiCYS-N and SiCYS-C coupled affinity columns; namely SiCYS CP, SiCYS-N CP and SiCYS-C CP, respectively. The protein bands with molecular weight of ~32, ~45, ~48 and ~52 kDa in SDS-PAGE were marked. (B) SiCYS CP, SiCYS-N CP and SiCYS-C CP were added with or without protease inhibitors, followed by zymographic analyses of protease activity. Lane 2 was added with 20 mM of PMSF; lane 3, 250 μM of E64; lane 4, 5 mM of iodoacetamide; lane 5, 5 mM of N-ethylmaleimide, lane 6, 100 μM of pepstatin A; lane 7, 25 mM of EDTA; while no any inhibitor was added in lane 1. The location of ~45 and ~52 kDa on gel was marked as well.