 | Acta Crystallographica Section F Acta Crystallographica Section F STRUCTURAL BIOLOGY COMMUNICATIONS |
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![[Figure 2]](no5090fig2mag.jpg)
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Figure 2
(a) Coomassie-stained SDS gel of purified proteins. Marker is shown in the left lanes (HiMark Protein marker; Life Technologies, USA; labelled in kDa). (b) Superposed size-exclusion chromatograms of proteins, normalized to 1 for the highest peak. The corynebacterial proteins run as monodisperse hexamers, while the M. tuberculosis FAS I chromatographic profile suggests conformational heterogeneity by additional aggregated and dimeric species. Slight shifts in elution volumes might result from different buffer conditions. Buffer conditions: M. tuberculosis FAS I, 0.1 M sodium phosphate pH 7.2, 0.1 M NaCl, 5 mM sodium malonate; C. ammoniagenes, 0.1 M sodium phosphate pH 6.5, 0.1 M NaCl, 5 mM sodium malonate; C. efficiens, 0.1 M bis-tris propane pH 6.8, 0.2 M NaCl, 10% glycerol. (c) Enzymatic activity of C. efficiens FAS I monitored by the consumption of NADPH at 334 nm. (d) Fluorescence-based thermal shift assay (Thermofluor assay) for buffer screening. (e) Thermal denaturation of protein monitored by CD spectroscopy. The melting points of the proteins were determined to be 45.4°C for M. tuberculosis FAS I, 44.6°C for C. ammoniagenes FAS I and 47.3°C for C. efficiens FAS I. (f) Crystals of C. efficiens FAS I. Colour code of figure: M. tuberculosis FAS I, black; C. ammoniagenes FAS I, red; C. efficiens FAS I, blue. |
![[Figure 2]](no5090fig2.jpg)
| STRUCTURAL BIOLOGY COMMUNICATIONS |
ISSN: 2053-230X
