| 摘要 |
The polyprenyl phosphate: N-acetylhexosamine-1-phosphate transferase family are glycosyl transferase enzymes of interest in the development of antibacterial treatments. The bacterial proteins WecA and MraY are exemplary transferases, each of which catalyzes the transfer of a specific hexosamine 1-P from a soluble UDP-hexosamine substrate to a polyprenyl phosphate carrier at the membrane surface. The present invention provides a generalizable, high throughput, one-pot assay for this type of enzymatic activity by incorporating a solid-liquid, bead-based separation system to selectively adsorb the highly hydrophobic products of the reaction. By judicious choice of radiolabeled UDP-hexosamine precursor, the assay format can be used to quantitate the products of diverse members of this transferase family as well as enzymes that catalyze the further modification of these transferase products. Thus, the use of this flexible assay format allows biochemical and enzymologic analysis of many such membrane bound transferases.
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