Rapid identification of DNA-binding proteins by mass spectrometry.

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 Biotechnol

journal_title

Nature biotechnology

authors

Nordhoff E,Krogsdam AM,Jorgensen HF,Kallipolitis BH,Clark BF,Roepstorff P,Kristiansen K

doi

10.1038/12873

subject

Has Abstract

pub_date

1999-09-01 00:00:00

pages

884-8

issue

9

eissn

1087-0156

issn

1546-1696

journal_volume

17

pub_type

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