 | Acta Crystallographica Section F Acta Crystallographica Section F STRUCTURAL BIOLOGY COMMUNICATIONS |
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![[Figure 1]](nj5302fig1mag.jpg)
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Figure 1
SDS–PAGE and SEC analyses of purified recombinant R. felis acetoacetyl-CoA reductase. (a) Acetoacetyl-CoA reductase eluted as a single peak (blue line) from a HiLoad 16/60 Superdex 200 pg SEC column. The SEC column was calibrated separately (gray line), and the molecular weights of calibration protein standards from a kit are indicated. The elution volume of R. felis acetoacetyl-CoA reductase falls between those of ovalbumin (75 kDa) and aldolase (158 kDa), suggesting a higher oligomeric state. SEC-purified acetoacetyl-CoA reductase was resolved on 12% SDS–PAGE visualized with Coomassie Blue (inset); the monomer runs as a single band (AcAc-CoA reductase) near the 25 kDa protein standard marker (lane M, labeled in kDa). A standard curve for SDS–PAGE (b) and a SEC calibration curve (c) were generated using the known protein standards. In (b), plotting the molecular weights (MW) of the protein standards versus their observed Rf (relative migration; black circles), the MW of the protein monomer was estimated to be 26 kDa (blue circle). In (c), the calibration curve was obtained by plotting Kav (the partition coefficient calculated using individual elution volumes) versus log relative molecular weight (Mr) of the protein standards. The Mr of R. felis acetoacetyl-CoA reductase was determined to be approximately 101 kDa (blue circle), indicating a tetrameric form, which is common in the short-chain dehydrogenase/reductase (SDR) family. |
![[Figure 1]](nj5302fig1.jpg)
| STRUCTURAL BIOLOGY COMMUNICATIONS |
ISSN: 2053-230X
