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. 2010 Sep 3;285(45):34401–34407. doi: 10.1074/jbc.M110.160515

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — PRA formation assay is dependent on TrpD, IlvA, PRPP and threonine. A standard assay for PRA formation included threonine, IlvA, PRPP, TrpD, cofactors, [¹⁴C]glycine, and PurD, as described under “Experimental Procedures.” PRA formation was determined as a function of [¹⁴C]GAR production. Lane 1, all reaction components present; lane 2, IlvA omitted; lane 3, TrpD omitted; lane 4, threonine omitted; lane 5, PRPP omitted. A, quantification of [¹⁴C]GAR produced under each condition, using [¹⁴C]glycine standards of known concentrations. Error bars represent the S.D. of two replicates. B, representative phosphorimage of [¹⁴C]GAR spots after separation from [¹⁴C]glycine by thin-layer chromatography.