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Title: Mass spectrometric characterization of sequence-specific complexes of DNA and transcription factor PU.1 DNA binding domain

Abstract

Electrospray ionization mass spectrometry (ESI-MS) has been used to study the noncovalent interaction of the 13.5-kDa DNA binding domain of PU.1 (PU.1-DBD) with specific double-stranded DNA (dsDNA) target molecules. Mixtures of PU.1-DBD protein and wildtype target DNA sequence yielded ESI-MS spectra showing only protein-dsDNA complex ions of 1:1 stoichiometry and free dsDNA. When PU.1-DBD protein, wild type target DNA, and a mutant target DNA lacking the consensus sequence were mixed, only the 1:1 complex with the wild-type DNA was observed, consistent with gel electrophoresis mobility shift assay results, demonstrating the observation of sequence-specific protein-dsDNA complexes using ESI-MS. 22 refs., 5 figs., 1 tab.

Authors:
; ;  [1]
  1. Pacific Northwest National Lab., Richland, WA (United States) [and others
Publication Date:
OSTI Identifier:
508281
DOE Contract Number:  
AC06-76RL01830
Resource Type:
Journal Article
Journal Name:
Analytical Biochemistry
Additional Journal Information:
Journal Volume: 239; Journal Issue: 1; Other Information: PBD: 15 Jul 1996
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA; MASS SPECTRA; PROTEINS; ELECTROPHORESIS; MOLECULAR STRUCTURE; TRANSCRIPTION FACTORS; MUTANTS; DNA SEQUENCING; BINDING ENERGY

Citation Formats

Cheng, Xueheng, Harms, A.C., and Bruce, J.E. Mass spectrometric characterization of sequence-specific complexes of DNA and transcription factor PU.1 DNA binding domain. United States: N. p., 1996. Web. doi:10.1006/abio.1996.0287.
Cheng, Xueheng, Harms, A.C., & Bruce, J.E. Mass spectrometric characterization of sequence-specific complexes of DNA and transcription factor PU.1 DNA binding domain. United States. doi:10.1006/abio.1996.0287.
Cheng, Xueheng, Harms, A.C., and Bruce, J.E. Mon . "Mass spectrometric characterization of sequence-specific complexes of DNA and transcription factor PU.1 DNA binding domain". United States. doi:10.1006/abio.1996.0287.
@article{osti_508281,
title = {Mass spectrometric characterization of sequence-specific complexes of DNA and transcription factor PU.1 DNA binding domain},
author = {Cheng, Xueheng and Harms, A.C. and Bruce, J.E.},
abstractNote = {Electrospray ionization mass spectrometry (ESI-MS) has been used to study the noncovalent interaction of the 13.5-kDa DNA binding domain of PU.1 (PU.1-DBD) with specific double-stranded DNA (dsDNA) target molecules. Mixtures of PU.1-DBD protein and wildtype target DNA sequence yielded ESI-MS spectra showing only protein-dsDNA complex ions of 1:1 stoichiometry and free dsDNA. When PU.1-DBD protein, wild type target DNA, and a mutant target DNA lacking the consensus sequence were mixed, only the 1:1 complex with the wild-type DNA was observed, consistent with gel electrophoresis mobility shift assay results, demonstrating the observation of sequence-specific protein-dsDNA complexes using ESI-MS. 22 refs., 5 figs., 1 tab.},
doi = {10.1006/abio.1996.0287},
journal = {Analytical Biochemistry},
number = 1,
volume = 239,
place = {United States},
year = {1996},
month = {7}
}