Toc75 | Protein TOC75-3, chloroplastic, POTRA domain 3
AS08 351 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A.thaliana, N. tabacum, P. patens, P. sativum, S. oleracea, T. salsuginea, some cross-Reactivity was observed for cyanobacteria including: Synechocystis, Synechococcus and Thermosynechococcus sp.
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Pisum sativum TOC75; Predicted POTRA Domain #3; Expressed and purified in E. coli using the Impact System from NEB. Peptide confirmed by MALDI. UniProt: Q43715
88 | 75 kDa (ocassionally a processing intermediate at 78 kDa is observed)
580 ng of Chl of Pisum sativum plants (10 day old) (2, 4) and 10 µg of combined envelopes of Pisum sativum 10 day-old (1,3) were separated on 15% SDS-PAGE and blotted 2h to PVDF. PVDF was blocked 1h with 3% non-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-Toc75 POTRA domain 3 antibodies AS08 351 (1:2000 and 1: 5000, 1h) and secondary donkey-anti-rabbit (1:20000, 1 h) antibody (HRP conjugated) in TBS-T containing 3% non fat milk powder. Antibody incubations were followed by washings in TBS-T. All steps were performed at RT with agitation. Blots were developed for 5 min with chemiluminescence detection reagent according to the manufacturers instructions. Exposure time was 600 seconds.
Ashita Dave and Barry D. Bruce (UTK), USA
OEP 75 or Toc75; Chloroplast outer envelope membrane protein from Pisum sativum (pea), Predicted to contain 3 POTRA domains at N-terminus. Believed to be the protein conducting channel of the Toc translocon and assembles as an 18 stranded ß-barrel (EMBO J. (1995) 14:11, 2436-2446); In Arabidopsis there are five members of this Family Toc75 (I-V), atToc75III is most closely related to psToc75. Additionally, it is structurally related to members of the bacterial surface antigen super-family including: OMA87; Outer membrane protein/protective antigen, (COG4775, COG4775) [Cell envelope biogenesis, outer membrane; YaeT; outer membrane protein assembly complex, (TIGR03303); FhaC; Hemolysin activation/secretion protein (COG2831) [Intracellular trafficking and secretion]
Vera-Estrella et al. (2014). Comparative 2D-DIGE analysis of salinity responsive microsomal proteins from leaves of salt-sensitive Arabidopsis thaliana and salt-tolerant Thellungiella salsuginea. J Proteomics. 2014 Jun 2. pii: S1874-3919(14)00288-7. doi: 10.1016/j.jprot.2014.05.018.
Hsueh et al. (2014). The chloroplast outer envelope protein P39 in Arabidopsis thaliana belongs to the Omp85 protein family. Proteins. 2014 Nov 17. doi: 10.1002/prot.24725.