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. 2022 Jun 13;204(7):e00078-22. doi: 10.1128/jb.00078-22

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Copyright © 2022 Heryakusuma et al.

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FIG 2 — Analysis for prosthetic groups of ANME2c-FsrII-6D. (A) UV-visible spectrum of FsrII. A 300-μL anaerobic solution of 84 μg of homogeneous ANME2c-FsrII-6D in 100 mM potassium phosphate buffer (pH 7) containing 250 mM NaCl was analyzed in a quartz cuvette with a 1-cm light path sealed with a no. 00000 rubber stopper (EPDM rubber stopper, WidgetCo, Houston, TX). (B) Reverse-phase HPLC analysis of a methanol-methylene chloride extract of ANME2c-FsrII-6D. (Main plot) Analysis of a 100-μL methanol-methylene chloride extract of 63.2 μg homogeneous protein. The peak corresponds to FAD. (Inset) Analysis of a 10-μL solution of 1 mM FAD and FMN in distilled water. (C) UV-visible spectrum of flavin cofactor extracted from ANME2c-FsrII-6D and resolved on a reverse-phase column as described for panel B.