Classical UDP-glucose 6-dehydrogenases (UGDHs; EC 18.104.22.168) catalyze the conversion of UDP-α-d-glucose (UDP-Glc) to the key metabolic precursor UDP-α-d-glucuronic acid (UDP-GlcA) and display specificity for UDP-Glc. The fundamental biochemical and structural study of the UGDH homolog CalS8 encoded by the calicheamicin biosynthetic gene is reported and represents one of the first studies of a UGDH homolog involved in secondary metabolism. The corresponding biochemical characterization of CalS8 reveals CalS8 as one of the first characterized base-permissive UGDH homologs with a >15-fold preference for TDP-Glc over UDP-Glc. The corresponding structure elucidations of apo-CalS8 and the CalS8·substrate·cofactor ternary complex (at 2.47 and 1.95 Å resolution, respectively) highlight a notably high degree of conservation between CalS8 and classical UGDHs where structural divergence within the intersubunit loop structure likely contributes to the CalS8 base permissivity. As such, this study begins to provide a putative blueprint for base specificity among sugar nucleotide-dependent dehydrogenases and, in conjunction with prior studies on the base specificity of the calicheamicin aminopentosyltransferase CalG4, provides growing support for the calicheamicin aminopentose pathway as a TDP-sugar-dependent process.
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This work was supported by National Institutes of Health Grants U01GM098248 (to G. N. P.), R01 CA84374 (to J. S. T.), and GM094585 (to A. J.) and the National Center for Advancing Translational Sciences (UL1TR000117).
Singh, Shanteri; Michalska, Karolina; Bigelow, Lance; Endres, Michael; Kharel, Madan K.; Babnigg, Gyorgy; Yennamalli, Ragothaman M.; Bingman, Craig A.; Joachimiak, Andrzej; Thorson, Jon S.; and Phillips, George N. Jr., "Structural Characterization of CalS8, a TDP-α-D-Glucose Dehydrogenase Involved in Calicheamicin Aminodideoxypentose Biosynthesis" (2015). Pharmaceutical Research and Innovation Faculty Publications. 1.