Fructosamine-3-kinase (FN3K) and the more recently discovered fructosamine-3-kinase related protein (FN3KRP) appear to protect proteins from nonenzymatic glycation. To elucidate the patterns of transcriptional regulation of these two genes, we performed in silico comparisons of their promoters along with real-time PCR assays of their expression in a variety of human tissues. Both promoters were TATA-less and CAAT-less, and contained several homologous CpG islands and Sp1 binding sites. The genes were expressed in all human tissues examined, with FN3K showing significantly higher levels in organs susceptible to nonenzymatic glycation and diabetic complications. Cultured fibroblasts treated with conditions mimicking the hormonal and biochemical profile of the diabetic state showed no changes in FN3K and FN3KRP expression relative to untreated cells. These data suggest that FN3K and FN3KRP act as protein repair enzymes and are expressed constitutively in human cells independently of some of the variables altered in the diabetic state.