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)
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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.