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. Author manuscript; available in PMC: 2017 Jan 24.
Published in final edited form as: Mol Biochem Parasitol. 2016 Jan 24;204(2):77–80. doi: 10.1016/j.molbiopara.2016.01.005

Table 1.

Oligonucleotides used as substrates for glucosyltransferase assay*

Substrate Sequence kcat (s−1) Km (μM) kcat/Km (μM−1 s−1)
WT2 5′-TTAGGGTTAGGGTTA-3′
3′-AATCCCAATCCCAAT-5′		 0.485 ± 0.022 0.519 ± 0.109 0.933
WT1 5′-TTAGGGTTAGGGTTA-3′
3′-AATCCCAATCCCAAT-5′		 0.451 ± 0.025 0.523 ± 0.137 0.862
WT3 5′-TTAGGGTTAGGGTTA-3′
3′-AATCCCAATCCCAAT-5′		 0.675 ± 0.037 0.519 ± 0.134 1.302
TTT 5′-TTTGGGTTTGGGTTT-3′
3′-AAACCCAAACCCAAA-5′		 0.703 ± 0.052 0.412 ± 0.149 1.705
ATT 5′-ATTGGGATTGGGATT-3′
3′-TAACCCTAACCCTAA-5′		 0.566 ± 0.034 0.267 ± 0.089 2.116
Random 5′-GTACGAGTCGAGTCA-3′
3′-CATGCTCAGCTCAGT-5′		 0.580 ± 0.021 0.313 ± 0.059 1.854
*

Bold T indicates the position of an hmU modification.

**

Glucosylation reactions were conducted with hmU DNA substrate concentration of 0, 0.195, 0.391, 0.781, 1.563, 3.125, 6.25, 12.5, and 25 μM and fixed enzyme and UDP-glucose concentrations of 18 μM and 1mM, respectively. UDP cleaved is plotted vs substrate concentration and nonlinear regression was performed to determine kinetic parameters.