Abstract:
:We report a protocol for the rapid identification of DNA-binding proteins. Immobilized DNA probes harboring a specific sequence motif are incubated with cell or nuclear extract. Proteins are analyzed directly off the solid support by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The determined molecular masses are often sufficient for identification. If not, the proteins are subjected to mass spectrometric peptide mapping followed by database searches. Apart from protein identification, the protocol also yields information on posttranslational modifications. The protocol was validated by the identification of known prokaryotic and eukaryotic DNA-binding proteins, and its use provided evidence that poly(ADP-ribose) polymerase exhibits DNA sequence-specific binding to DNA.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Nordhoff E,Krogsdam AM,Jorgensen HF,Kallipolitis BH,Clark BF,Roepstorff P,Kristiansen Kdoi
10.1038/12873subject
Has Abstractpub_date
1999-09-01 00:00:00pages
884-8issue
9eissn
1087-0156issn
1546-1696journal_volume
17pub_type
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