Sucrose protects cell wall invertase but not vacuolar invertase against proteinaceous inhibitors

Vacuolar (VI) and cell wall invertases (CWI) of higher plants can be inactivated in vitro and, possibly, in vivo by proteinaceous inhibitors. The respective mechanisms have not yet been compared. Therefore, partially purified CWI from transformed tobacco cells and VI from tomato fruit were preincubated with invertase-inhibitor fractions isolated from the same tissues. Both inhibitors were able to inhibit both invertases. However, VI was fully inhibited within less than 1 min by both inhibitors, whereas inactivation of CWI was much slower. Furthermore, CWI, but not VI, was strongly protected against inhibition by sucrose. A polyclonal antiserum directed against the tobacco inhibitor (I(NT)) cross-reacted with a 19 kDa polypeptide in the partially purified tomato inhibitor (I(LE)) fraction. The results indicate that I(NT) and I(LE)have similar structural properties, whereas the mechanism of inactivation is clearly different for CWI and VI.