Labeling of fusion proteins of O6-alkylguanine-DNA alkyltransferase with small molecules in vivo and in vitro

The in vivo and in vitro labeling of fusion proteins with synthetic molecules capable of probing and controlling protein function has the potential to become an important method in functional genomics and proteornics. We have recently introduced ail approach for the specific labeling of fusion proteins, which is based on the generation of fusion proteins with the human DNA repair protein O-6-alkylguanine-DNA alkyltransferase (hAGT) and the irreversible reaction of hAGT with O-6-benzylguanine derivatives. Here, we report optimized protocols for the synthesis of O-6-benzylguanine derivatives and the use of such derivatives for the labelling of different hAGT fusion Proteins in vivo and in vitro. (C) 2003 Elsevier Inc. All rights reserved.