A method of assaying for glycosyltransferase activity in a sample. In a first step, a sample is reacted with a first sugar donor and an acceptor substrate to produce a transferase product. The first sugar donor and acceptor substrate are selected such that the sugar from the first sugar donor is capable of being transferred to the acceptor substrate in the presence of the glycosyl-transferase to be assayed. In a second step, the transferase product is reacted with a second sugar donor having a sugar which is labelled with a labelling agent and an enzyme which is capable of transferring the sugar from the second sugar donor to the transferase product to produce a labelled transferase product and which has a higher affinity for the glycosyltransferase product compared to the affinity of the glycosyltransferase for the acceptor substrate. The labelling agent activity of the labelled transferase product or unreacted second sugar donor is assayed to determine transferase activity in the sample. A kit for assaying for glycosyltransferase activity in a sample is also described.
METHOD FOR MEASURING GLYCOSYLTRANSFERASE ACTIVITY
DATTI, Alessandro
1992
Abstract
A method of assaying for glycosyltransferase activity in a sample. In a first step, a sample is reacted with a first sugar donor and an acceptor substrate to produce a transferase product. The first sugar donor and acceptor substrate are selected such that the sugar from the first sugar donor is capable of being transferred to the acceptor substrate in the presence of the glycosyl-transferase to be assayed. In a second step, the transferase product is reacted with a second sugar donor having a sugar which is labelled with a labelling agent and an enzyme which is capable of transferring the sugar from the second sugar donor to the transferase product to produce a labelled transferase product and which has a higher affinity for the glycosyltransferase product compared to the affinity of the glycosyltransferase for the acceptor substrate. The labelling agent activity of the labelled transferase product or unreacted second sugar donor is assayed to determine transferase activity in the sample. A kit for assaying for glycosyltransferase activity in a sample is also described.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
