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NF-KB protein purification from bovine spleen: Nucleotide stimulation and binding site specificity

Lenardo, Michael J. and Kuang, Anna and Gifford, Ann and Baltimore, David (1988) NF-KB protein purification from bovine spleen: Nucleotide stimulation and binding site specificity. Proceedings of the National Academy of Sciences of the United States of America, 85 (23). pp. 8825-8829. ISSN 0027-8424. PMCID PMC282599. doi:10.1073/pnas.85.23.8825.

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The activity of the enhancer for the κ immunoglobulin light chain gene critically depends on the presence in the nucleus of the NF-κB protein. We purified NF-κB over 50,000-fold and identified two protein species, 42 and 44 kDa, that could be eluted and renatured from a sodium dodecyl sulfate/polyacrylamide gel to give specific DNA-binding activity. Binding of the purified bovine NF-κB as well as that from human and murine B- or T-lymphoid cell extracts was dramatically stimulated by nucleoside triphosphates. This effect distinguished NF-κB from a related factor, H2-TF1. Purified NF-κB interacted efficiently with regulatory sequences that function during either B- or T-lymphocyte activation, including the human immunodeficiency virus enhancer and a NF-κB binding site we detected in the interleukin 2 enhancer.

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Baltimore, David0000-0001-8723-8190
Additional Information:© 1988 by the National Academy of Sciences. Contributed by David Baltimore, August 10, 1988. We thank Cornelis Murre, Patrick Baeuerle, Patrick McCaw, Louis Staudt, Mark Feinberg, Paul Matsudaira, and Paul Robbins for assistance in the purification and for nuclear extracts. We also thank Sharon Cross, Warren Leonard, and Harinder Singh for probes and David Weaver and Steve Smale for a critical review of the manuscript. M.J.L. was supported by a Clinical Investigator Award of the National Cancer Institute, and the work was funded by National Institutes of Health and American Cancer Society grants to D.B. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:κ immunoglobulin light chain gene; enhancer; interleukin 2; human immunodeficiency virus
Issue or Number:23
PubMed Central ID:PMC282599
Record Number:CaltechAUTHORS:LENpnas88
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8094
Deposited By: Tony Diaz
Deposited On:31 Jul 2007
Last Modified:08 Nov 2021 20:47

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