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. 2015 Jan 30;10(1):e0116750. doi: 10.1371/journal.pone.0116750

Figure 4. Oligomerization of the PHB domain.

Figure 4

(A) Size exclusion chromatography was performed with the PHB domain on a Superose6 column (GE Healthcare) and two elution peaks were detected at 14.6 and 17.5 ml, corresponding to a size of 313 kDa and 42.8 kDa respectively. (B) Blue native PAGE was performed with the PHB domain, three bands marked with red arrows could be detected corresponding to a monomer, a 14mer (313 kDa) and an oligomer larger than 670 kDa. (C) A colloidal Coomassie blue stained SDS—PAGE gel is shown. The PHB domain was further probed by cross-linking and applying the sample to SDS-PAGE. For cross-linked PHB only a band larger than 250 kDa is detected, for none cross-linked PHB domain bands corresponding to a monomer, a tetramer, an octamer and an oligomer larger than 250 kDa are be detected (indicated by blue arrows). The SEC column was calibrated using standard proteins (see S2 Fig.).