Fig. 5.
In vitro activation of apo‐yAnfDK using purified FeMo‐co. Preparations of apo‐yAnfDK with His10, Twin‐Strep (TS) or Single‐Strep (SS) purification tags at the N‐terminal end of AnfD were obtained from GF15 (expressing AnfHGHisDK), GF17 (expressing AnfHGTSDK) or GF19 (expressing AnfHGSSDK) respectively. Five µl FeMo‐co isolated from A. vinelandii was added to 6 µM of GF17 apo‐yAnfDK or 12 µM of GF15 or GF19 apo‐yAnfDK. Positive control reactions were carried with 11 µM of A. vinelandii apo‐NifDK. The activity of reconstituted yAnfDK was determined by the acetylene reduction assay after addition of AvNifH and ATP‐regenerating mixture. Data represent mean ± SD (n = 6) for each yAnfDK.