Figure 2.

In vitro hydrolysis of HsEg5_1-367 motor domain by s-ZPrC

Recombinant soluble s-ZPrC (27 kDa) was incubated with purified HsEg5 motor domain (42 kDa) for 24 h. s-ZPrC exhibited a low level of self-cleavage after 24 h marked primarily by a 23 kDa fragment. After 24 h, HsEg5 was cut into 21.7 kDa N-terminal (red arrow) and 20.4 kDa C-terminal (cyan arrow) fragments. In turn, the 20.4 kDa fragment was incompletely cut into 9.7 kDa and 10.7 kDa fragments (asterisk). The 20.4 kDa fragment was excised for sequencing to identify the primary s-ZPrC cleavage site (Figure 3).