A study of the envelope proteins of the BHK-21 cell nucleus

Wilson, Virginia S (1983) A study of the envelope proteins of the BHK-21 cell nucleus. MSc(R) thesis, University of Glasgow.

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Abstract

Fractions of BHK cells were examined in both the normal C13 line and after transformation with Polyoma virus (PyY). Protein profiles of whole nuclei, nuclear envelope, pore complex lamina, nuclear membrane and rough endoplasmic reticulum were compared and the carbohydrates detected by their affinity for [125 I] - labelled lectins. Carbohydrates were found in all these fractions with minor differences in their relative proportions after transformation, and a loss of all affinity for RCA. Similarities in the affinities for WGA and lentil lectin were seen in nuclear membrane and rough endoplasmic reticulum suggesting that these membranes may have a common origin. A large proportion of the nuclear envelope carbo-hydrate was retained by the lamina after its separation from the membrane. A specific interaction between transmembrane glycoproteins and the lamina is proposed. The interaction of lectins with whole nuclei was examined. This revealed some changes in the affinities and number of binding sites/nucleus after transformation. The topography of the nuclear envelope was examined by comparison of the iodination profiles produced by soluble and immobilised lactoperoxidase. Nuclei were examined before and after removal of their membranes with Triton-X100. In the absence of the membrane, the lamina topography was examined by comparative iodinations after contraction and expansion in various concentrations of Ca2+. In whole nuclei the iodination profile of immobilised lacto-peroxidase was very restricted, with heavy iodination in only one band of Mr 52,000. This was reduced by Triton-X100 extraction ii which produced a concomitant increase in the labelling of a protein of Mr 63,000. This may indicate that it lies below the membrane on the lamina surface. Contracted nuclei from which the membrane had been removed presented a similar range of proteins to both soluble and immobilised lactoperoxidase indicating a very close association between the proteins. After expansion, the iodination profiles of both agents were extended, suggesting that the proteins had undergone dissociation.

Item Type: Thesis (MSc(R))
Qualification Level: Masters
Additional Information: Adviser: J G Beeley
Keywords: Cellular biology
Date of Award: 1983
Depositing User: Enlighten Team
Unique ID: glathesis:1983-73356
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 14 Jun 2019 08:56
Last Modified: 14 Jun 2019 08:56
URI: http://theses.gla.ac.uk/id/eprint/73356

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