Cell-free expression of human glucosamine 6-phosphate N-acetyltransferase (HsGNA1) for inhibitor screening

► Functional human GNA1 and GNA1-sGFP were highly expressed by cell-free system. ► Inhibition assay with glucose-6-phosphate was performed and IC50 was calculated. ► Binding sites of glucose-6-phosphate to GNA1 were identified by simulated annealing. ► This strategy dramatically reduces the timeline for GNA1 analysis and screening.