Skip to main content
. Author manuscript; available in PMC: 2018 Nov 29.
Published in final edited form as: Methods Enzymol. 2018 Jun 1;606:485–522. doi: 10.1016/bs.mie.2018.04.014

Fig. 8.

Fig. 8

Isolation of 3′,8-cH2GTP. (A) Chromatograms of 3′,8-cH2GTP purification on QAE Sephadex A25 (left) and DEAE Sepharose FF (right) columns. The gradient of NH4HCO3 (pH 9) buffer for each chromatography is indicated by dotted lines. The amount of 3′,8-cH2GTP is quantified by either UV absorbance (gray dashed lines) at 322nm or MoaC assay (black solid lines). The amount of GTP is quantified by UV absorbance at 256nm after subtraction of the contribution from 3′,8-cH2GTP based on its extinction coefficient at 256nm (black dashed lines). (B) Overall scheme for the 3′,8-cH2GTP isolation. Panel (A) is reprinted with permission from Hover, B. M., Loksztejn, A., Ribeiro, A. A., & Yokoyama, K. (2013). Identification of a cyclic nucleotide as a cryptic intermediate in molybdenum cofactor biosynthesis. Journal of the American Chemical Society, 135(18), 7019–7032. doi:10.1021/ja401781t. Copyright 2013 American Chemical Society.