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. 2021 Jul 19;118(30):e2102318118. doi: 10.1073/pnas.2102318118

Fig. 7.

Fig. 7.

[3Fe-4S] is essential for the catalytic activity of BlsK. (A) HPLC analysis of BlsK-catalyzed reaction: (i) using BlsK purified under aerobic conditions; (ii) with BlsK purified under anaerobic conditions; and (iii) BlsK purified under anaerobic conditions with the [3Fe-4S] further reconstituted. (B) EDTA abolished the activity of BlsK: (i) without EDTA as control; (ii) BlsK treated with EDTA; and (iii) BlsK treated with EDTA following by the reconstitution of iron–sulfur cluster in vitro. (C) UV-visible analysis of BlsK treated with EDTA. (D) HPLC analysis of the activity of BlsK mutant proteins: (i) wild-type BlsK, and (ii–iv) represent C236S, C253S, and C259S mutant proteins, respectively. The concentration of BlsK in the reaction system was 10 μM. Enzyme assays in A and B were carried out under anaerobic conditions.