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Fig. 3 | BMC Plant Biology

Fig. 3

From: Chloroplast PetD protein: evidence for SRP/Alb3-dependent insertion into the thylakoid membrane

Fig. 3

Import of the PetD protein into isolated pea thylakoid. a Insertion of radiolabeled PetD in the presence of thylakoid membrane, supernatant (S) and membrane pellet (M) after fractionation, the total translation mixture is marked TMi, b TMi- insertion of the translated PetD in the presence of thylakoid membrane and stromal protein fractions, and the S-supernatant and M-membrane pellet after fractionation. c Chaotropic extraction of inserted PetD by NaSCN, TMi-fraction after PetD insertion (control); S-supernatant and M-membrane pellet after fractionation. d Insertion of PetD in the presence of thylakoid membrane and Ab against ALB3 protein (denoted with “+”). TMi-fraction after PetD insertion (control); S-supernatant and M-membrane pellet after fractionation. e Insertion of translated PetD in the presence of thylakoid membrane and Ab against cpSECY protein (denoted with “+”). TMi-fraction after PetD insertion (control); M -membrane pellet; S-supernatant after fractionation. f Western blot analysis of chaotropic extraction of PetA subunit from thylakoids membrane with inserted PetD. TMi-fraction after PetD insertion (control); S-supernatant and M-membrane pellet after fractionation. NaSCN or NaBr was used as the chaotropic agent [33]. The experiments were repeated at least twice. Each lane was loaded with 15 μg of total protein aliquot. Membranes were incubated with appropriate Ab before insertion started and washed twice. Fig’s a − e present autoradiograph analysis. MW of radiolabeled product was determined by MALDI-TOF

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