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. Author manuscript; available in PMC: 2015 Feb 6.
Published in final edited form as: J Mol Biol. 2014 Feb 6;426(3):570–585. doi: 10.1016/j.jmb.2013.10.025

Fig 3.

Fig 3

HoloCcmE co-purifies with CcmF in the absence of CcmGH. (A) Coomassie blue staining of TALON-purified proteins from ΔFGH, ΔEGH, ΔGH, and pSysI backgrounds showing purified 54-kDa CcmF. (B) Anti-CcmF immunoblot of TALON-purified proteins showing 54-kDa CcmF. (C) Anti-CcmH immunoblot of TALON-purified proteins showing 34 kDa CcmH. (D) Heme staining of TALON-purified proteins showing free heme (CcmF b-heme) and co-purified 20-kDa holoCcmE. (E) Anti-CcmE immunoblot of TALON-purified proteins showing co-purified 20-kDa CcmE. For (A)-(E), 6 ug purified protein was analyzed. (F) Heme staining of DDM-solubilized membrane fractions from ΔFGH, ΔEGH, ΔGH, and pSysI backgrounds showing 20 kDa holoCcmE. (G) Anti-CcmE immunoblot of DDM-solubilized membrane fractions showing 20 kDa CcmE. For (F) and (G), 70 ug total protein was analyzed. (H) Quantification of the results of heme staining (holoCcmE) and anti-CcmE immunoreactivity (total CcmE) from TALON-purified fractions from three independent experiments. (I) Quantification of the results of heme staining and anti-CcmE immunoreactivity from DDM-solubilized membrane fractions from three independent experiments. For (H) and (I), percent holoCcmE and total CcmE is relative to ΔGH, which has been set at 100%. Error bars denote SD.