Xenopus egg extracts: a model system to study proprotein convertases.

Abstract

The Xenopus egg extract translation system has proved an ideal tool with which to study the biosynthesis of the prohormone convertases. It provides a robust coupled translation/translocation system capable of efficient translocation of any protein containing an N-terminal signal sequence into the lumen of its microsomal membranes, with cotranslational cleavage of the signal peptide. Its main advantage over rival in vitro translation systems is that it will also carry out posttranslational modification of proteins, such as N-glycosylation, and, in the case of the proprotein convertases, support autocatalytic proregion removal. The egg extract also contains an endogenous, acidic pH optimum enzyme activity, suggestive of a proprotein convertase, that can undertake limited proteolysis of precursors containing multibasic processing sites.