Separation of Nuclear Pore Complex Lamina Proteins by Hydrophobic Chromatography
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Authors
Quinn, Kathleen
Issue Date
1983
Type
Thesis
Language
en_US
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Abstract
These experiments represent the first attempts to separate three polypeptides found in the nuclear pore complex lamina. The nuclear pore complex lamina (PCL) is a structural component of eucaryotic nuclei, lying on the nucleoplasmic face of the inner nuclear membrane. The PCL is composed mainly of fibrous proteins. In the bovine liver, the three major proteins constituting the PCL have molecular weights of 78,000, 73,000, and 63,000 daltons, as determined by previous studies. Recent work by others, resulting in a method of solubilizing the bovine liver PCL in 2% deoxycholate (DOC) and 3% 2-B-mercaptoethanol (2-ME), was the starting point for this study. This set of experiments attempted to separate the three major lamins found in bovine liver PCL. Based on suspected hydrophobicity differences of the lamins, hydrophobic column chromatography was the method utilized to separate the proteins. Removal of the DOC from the solubilized PCL mixture was necessary before chromatography could be carried out. DOC was removed in this study by dialysis. DOC concentration was shown to decrease from 0.2% to 0.74 x lO-4% without protein precipitation. Lamin separation was attempted once excess DOC was removed. Use of a series of agarose resins indicated that the three major lamins were hydrophobic but did not offer a viable means of protein separation. A Phenyl Sepharose resin, in contrast, produced results which suggest that further studies using stepwise elution of this column may well result in clean separation of lamin B.
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iv, 40 p.
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