Use of biotinylated NAD to label and purify ADP-ribosylated proteins.

摘要: Abstract Biotin- or digoxigenin-conjugated NAD has been used successfully to label EF-2 by diphtheria toxin,3 an α subunit of G protein pertussis toxin,4 and poly(ADP-ribose) synthase through auto-poly(ADP-ribosyl)ation (J. Zhang, unpublished result, 1996). It is likely that many other ADP-ribosyltransferases are capable using modified as substrates. Compared radioactive labeling, biotinylation several advantages. Commercially available precursors make synthesis biotinylated simple economic. No extensive purification the product required. Because can be separated from readily, there no dilution, in contrast [32P]NAD, which only a small proportion molecules radioactive. Once purified, stored for long time without decay (unlike NAD, does decay). Most importantly, system described here may afford efficient means purifying identifying ADP-ribosylated proteins. Biotinylated situ labeling study cellular localization tissue distribution