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Fig. 4 | BMC Microbiology

Fig. 4

From: Bacterial microcompartments for isethionate desulfonationĀ in the taurine-degrading human-gut bacterium Bilophila wadsworthia

Fig. 4

Sucrose gradient purification of BMCs from cell-free extracts of B. wadsworthia.Ā (A) Photograph of a sucrose gradient after centrifugation. The numbers mark the fractions that were collected. Fraction 2 contained most of the soluble cytosolic proteins, fraction 3 broken BMCs, while fraction 4 contained intact BMCs (see C). Fraction 10 contained also the centrifugation pellet; it was resuspended in fraction-10 solution (0.5 ml) and included in the SDS-PAGE analysis. (B) Nine excerpts of negative-stained TEM images of fraction 4, each showing sections where microcompartment-like structures were observed. (C) Double-stained (Coomassie and silver) SDS-PAGE gel of the gradient fractions (lanes from left to right: size marker (M), fractions 1 through 8, fraction 10). The upper red box marks the molecular weight of the signature enzyme of the BMC, isethionate sulfite-lyase (IslA) at approximately 94Ā kDa. The strong band at 40Ā kDa correspond to SarD (40.8Ā kDa) and Ald (39.8Ā kDa). The purple boxes mark bands that were identified by proteomic analysis as BMC shell proteins for fractions 4 and fraction 3 (see main text), which were visible only through silver staining. Fraction 9 was not included in the analysis as it contained only an insignificant amount of protein. M, protein molecular mass markers

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