Influence of the excision shock on the protein metabolism ofVicia faba L. meristematic root cells

UsingVicia faba root meristems we have shown that protein synthesis was dramatically changed after excision. The amino-acid incorporation dropped to 13% of the level in the unexcised control. This downshift was a direct consequence of the breakdown of polysomes which are converted into monosomes. In order to perform an analysis of the protein pattern by two-dimensional gel electrophoresis, endogenous proteolytic activity, which is high in broad bean root, had to be inhibited. Therefore, several protease inhibitors were tested and a very efficient inhibitor pool was obtained which could be used during the preparation of meristematic cell extracts. Protein-pattern analysis showed important differences between the unexcised control and excised apices. The number of proteins synthesized after excision droped from 250 in the control to 80, as a consequence of polysome breakdown. Futhermore, we present evidence that new and apparently specific proteins are synthesized in response to this excision shock.Abbreviations NEM N-ethylmaleimide - PMSF phenylmethylsulfonyl fluoride - TLCK N-tosyl-L-lysin chloromethyl ketone - TPCK N-tosyl-L-phenylalanine chloromethyl ketone