Proteolytlc digestion of the 2 4 WO-M, precursor and the 11 OOO-Mr transported membrane protein Preparation nf gel slices containing the bands of interest Preparative gel electrophoreses of either supernatants from trlchloroac-tic acrd preclpltatron of translation products in vitro 1200 pl t r a n s l a t i o n mixi or chloroplast membranes after transpurt 1400 pl translation supernatant were transported) W e r e performed o n 10-15(. gradient polyacrylamide gels. The bands were localized by added or internal menbrane pratems and stainrng. I:,. addlflon, t'le radloactlvrty Of the selected slices was compared with their nerghbourrng bands; in the j a e e of the gel from transport, no contaminating radioactivity was present. according to Wieland and Geargopulos (291. After drying, the plates were treated w l t h enhance spray (New Engl.ind Nuclear) and exposed on Du Pont Cronex 2 f i l m . Proteolytlc digestion with Staphylococcus aureus protease YBGel slices of 1 cm lengtii 11/15 of a preparative gel) were prerncubated for 30 mrn at room temperature i n 0.1 M NH HC03 and 0.0016% SDS. After homogenization In 0.9 ml of the same but fresh medlum, the protelns were eluted by rncubatlon for 1 h aL 55OC. The extraction was repeated twice and the combined supernatants were lyaphrlired. S . aureus protease. ThP prntelni were dlgested and separated
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