L.I. Nagore and R.J. Nadeau contributed equally to the study.
Purification and characterization of transcription factors†
Article first published online: 7 JUL 2013
© 2013 Wiley Periodicals, Inc.
Mass Spectrometry Reviews
Volume 32, Issue 5, pages 386–398, September/October 2013
How to Cite
Nagore, L.I., Nadeau, R.J., Guo, Q., Jadhav, Y.L.A., Jarrett, H.W. and Haskins, W.E. (2013), Purification and characterization of transcription factors. Mass Spectrom. Rev., 32: 386–398. doi: 10.1002/mas.21369
- Issue published online: 23 AUG 2013
- Article first published online: 7 JUL 2013
- Manuscript Revised: 19 NOV 2012
- Manuscript Accepted: 19 NOV 2012
- Manuscript Received: 31 JAN 2012
- NIH. Grant Number: G12MD007591
- transcription factor;
- capillary liquid chromatography;
- tandem mass spectrometry;
- protein database searching;
- response element;
Transcription factors (TFs) are essential for the expression of all proteins, including those involved in human health and disease. However, TFs are resistant to proteomic characterization because they are frequently masked by more abundant proteins due to the limited dynamic range of capillary liquid chromatography-tandem mass spectrometry and protein database searching. Purification methods, particularly strategies that exploit the high affinity of TFs for DNA response elements (REs) on gene promoters, can enrich TFs prior to proteomic analysis to improve dynamic range and penetrance of the TF proteome. For example, trapping of TF complexes specific for particular REs has been achieved by recovering the element DNA–protein complex on solid supports. Additional methods for improving dynamic range include two- and three-dimensional gel electrophoresis incorporating electrophoretic mobility shift assays and Southwestern blotting for detection. Here we review methods for TF purification and characterization. We fully expect that future investigations will apply these and other methods to illuminate this important but challenging proteome. © 2013 Wiley Periodicals, Inc. Rapid Commun. Mass Spectrom. 32: 386–398, 2013.