Shinde U, Inouye M
Department of Biochemistry Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway 08854, U.S.A.
J Mol Biol. 1995 Mar 31;247(3):390-5. doi: 10.1006/jmbi.1994.0147.
Subtilisin is synthesized with an N-terminal propeptide which has been demonstrated to function as an intramolecular chaperone that is only essential for the folding of the active enzyme. After folding, the propeptide is removed via an intramolecular autoprocessing mechanism. This mechanism is blocked when His64, a member of the catalytic triad is substituted with Ala. However, an additional mutation in the propeptide substituting Glu-2 with His was able to suppress the His64Ala mutation, allowing autoprocessing of the propeptide. This suppression is considered to be due to a "substrate assisted catalysis" mechanism and demonstrates that the cleavage to the subtilisin propeptide is an autocatalytic process.
枯草杆菌蛋白酶合成时带有一个N端前肽,该前肽已被证明作为分子内伴侣发挥作用,对活性酶的折叠至关重要。折叠后,前肽通过分子内自加工机制被去除。当催化三联体成员之一的His64被Ala取代时,该机制被阻断。然而,前肽中Glu-2被His取代的额外突变能够抑制His64Ala突变,使前肽能够进行自加工。这种抑制被认为是由于“底物辅助催化”机制,表明枯草杆菌蛋白酶前肽的切割是一个自催化过程。