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Purification and analysis of RNA polymerase II transcription factors by using wheat germ agglutinin affinity chromatography


, : Purification and analysis of RNA polymerase II transcription factors by using wheat germ agglutinin affinity chromatography. Proceedings of the National Academy of Sciences of the United States of America 86(6): 1781-1785

We recently found that many RNA polymerase II transcription factors are modified with N-acetylglucosamine residues. These sugar moieties confer upon transcription factors an ability to bind the lectin wheat germ agglutinin. We have taken advantage of this interaction to devise a purification procedure for the "GC-box" binding transcription factor Sp1. Crude nuclear extracts are first subjected to wheat germ agglutinin affinity chromatography and then subjected to sequence-specific DNA affinity chromatography. The Sp1 protein purified by this procedure is at least 95% pure, and the overall recovery is greater than 80%. In addition to yielding larger quantities of Sp1 than conventional schemes, the new purification procedure is also simpler and more rapid. We show that wheat germ agglutinin affinity chromatography can also be used to purify the glycosylated forms of the CCAAT-binding transcription factor. Thus, wheat germ agglutinin affinity chromatography may aid the purification of other transcription factors that bear N-acetylglucosamine residues. Furthermore, the ability to separate glycosylated forms of transcription factors from their unglycosylated counterparts by wheat germ agglutinin affinity chromatography should facilitate investigations into the role of N-acetylglucosamine residues in the functioning of transcription factor proteins.

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PMID: 2648381


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Schmidt, H.; Omran, H.; Sommer, T.; Steffens, M.; Illien, S.; Boes, M.; Schild, H.; Luederitz, B., 2001: Incidence of cerebral embolism in patients with atrial fibrillation and atrial thrombi and/or spontaneous echo contrast A prospective and serial study. Journal of the American College of Cardiology 37(2 Supplement A): 103A-104A, February

Rodhe, Henning, 1992: Modeling biogeochemical cycles. Butcher, Samuel S [Editor], Charlson, Robert J [Editor], Orians, Gordon H [Editor], Wolfe, G V [Editor] International Geophysics Series; Global biogeochemical cycles : 55-72

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McKee Brent A., 1989: Uranium-series radionuclides in the Mississippi River. Open-File Report - U S

Marshall, S.; Bauer, J.; Isenring, E., 2014: The consequences of malnutrition following discharge from rehabilitation to the community: a systematic review of current evidence in older adults. The prevalence of malnutrition in the rehabilitation setting is estimated to be 30-50%, with older adults at higher nutritional risk. Malnutrition also exists in the community setting, where 10-30% of adults are malnourished; however, the relation...

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Lobl, I.; Okali, I., 1969: List of the types in the collection of the Slovak National Museum: Coleoptera. Sbornik SIovenskeho Narodneho Muzea, 15(2): 39-66

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