Effect of dithiothreitol during isolation of nuclear matrix preparations from rat liver and Zajdela's hepatoma on their protein composition and phosphorylation

1991 ◽  
Vol 111 (4) ◽  
pp. 523-527
Author(s):  
D. G. Mal'dov
Keyword(s):  
Rat Liver ◽  
Nuclear Matrix ◽  
Protein Composition ◽  
Zajdela's Hepatoma

scholarly journals Epithelial structure revealed by chemical dissection and unembedded electron microscopy.

1984 ◽  
Vol 99 (1) ◽  
pp. 203s-208s ◽  
Author(s):  
E G Fey ◽  
D G Capco ◽  
G Krochmalnic ◽  
S Penman
Keyword(s):  
Nuclear Matrix ◽  
Intermediate Filament ◽  
Protein Composition ◽  
Intact Cells ◽  
Thin Sections ◽  
Transmission Electron ◽  
Kidney Epithelial Cell ◽  
Epithelial Structure ◽  
Triton X ◽  
Whole Mounts

Cytoskeletal structures obtained after extraction of Madin-Darby canine kidney epithelial cell monolayers with Triton X-100 were examined in transmission electron micrographs of cell whole mounts and unembedded thick sections. The cytoskeleton, an ordered structure consisting of a peripheral plasma lamina, a complex network of filaments, and chromatin-containing nuclei, was revealed after extraction of intact cells with a nearly physiological buffer containing Triton X-100. The cytoskeleton was further fractionated by extraction with (NH4)2SO4, which left a structure enriched in intermediate filaments and desmosomes around the nuclei. A further digestion with nuclease and elution with (NH4)2SO4 removed the chromatin. The stable structure that remained after this procedure retained much of the epithelial morphology and contained essentially all of the cytokeratin filaments and desmosomes and the chromatin-depleted nuclear matrices. This structural network may serve as a scaffold for epithelial organization. The cytoskeleton and the underlying nuclear matrix intermediate filament scaffold, when examined in both conventional embedded thin sections and in unembedded whole mounts and thick sections, showed the retention of many of the detailed morphological aspects of the intact cells, which suggests a structural continuum linking the nuclear matrix, the intermediate filament network, and the intercellular desmosomal junctions. Most importantly, the protein composition of each of the four fractions obtained by this sequential procedure was essentially unique. Thus, the proteins constituting the soluble fraction, the cytoskeleton, the chromatin fraction, and the underlying nuclear matrix-intermediate filament scaffold are biochemically distinct.

Considerations in the isolation of rat liver nuclear matrix, nuclear envelope, and pore complex lamina

1981 ◽  
Vol 132 (1) ◽  
pp. 105-123 ◽  
Author(s):  
Scott H. Kaufmann ◽  
Donald S. Coffey ◽  
Joel H. Shaper
Keyword(s):  
Nuclear Envelope ◽  
Rat Liver ◽  
Nuclear Matrix

The Protein Composition of the Hepatocyte Nuclear Matrix Is Differentiation-Stage Specific

IUBMB Life ◽  
2000 ◽  
Vol 49 (6) ◽  
pp. 511-517 ◽  
Author(s):  
Svetlana Ivanovi &#39 -Mati &#39 , Svletlana
Keyword(s):  
Nuclear Matrix ◽  
Protein Composition ◽  
Differentiation Stage

Protein composition of nuclear matrix preparations from HeLa cells: an immunochemical approach

1986 ◽  
Vol 80 (1) ◽  
pp. 103-122
Author(s):  
R. Verheijen ◽  
H. Kuijpers ◽  
P. Vooijs ◽  
W. van Venrooij ◽  
F. Ramaekers
Keyword(s):  
Nuclear Matrix ◽  
Connective Tissue Diseases ◽  
Protein Composition ◽  
Cytoskeletal Proteins ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Core Proteins ◽  
Nuclear Rna ◽  
Hela S3 ◽  
Protein Components

Procedures for the isolation of HeLa S3 nuclear matrices were re-examined with special emphasis on the use of various nucleases and detergents as well as on the ionic strength of the final salt extraction. The protein composition of the resulting nuclear matrix preparations was analysed by one- and two-dimensional gel electrophoresis and found to be extremely reproducible. By means of co-electrophoresis several typical cytoskeletal proteins (actin, vimentin and cytokeratins) and heterogeneous nuclear RNA (hnRNA)-associated core proteins (hnRNP) were shown to be present in such nuclear matrix preparations. The nature of some other protein components was elucidated using two-dimensional immunoblotting and immunofluorescence. For this purpose mouse monoclonal antibodies to cytoskeletal components (vimentin, cytokeratins), small nuclear RNP (70 X 10(3) Mr protein of U1-RNP), hnRNP (C1/C2) and the pore-complex lamina (lamins A, B and C) were used next to human autoimmune sera obtained from patients with connective tissue diseases and directed against the residual nucleoli and the internal fibrillar mass. These antibodies enabled us to identify a number of proteins present specifically in the nuclear matrix and to show that part of the cytoskeletal proteins are still present in the isolated structures.

DNA Sequences Nonrandomly Distributed in Respect to Rat Liver Nuclear Matrix

1990 ◽  
pp. 333-336
Author(s):  
J. Rzeszowska-Wolny ◽  
J. Lanuszewska ◽  
J. Rogolinski
Keyword(s):  
Rat Liver ◽  
Dna Sequences ◽  
Nuclear Matrix

scholarly journals Interactions of the matrix attachment region of DNA with the matrix proteins from the copper preincubated liver nuclei.

1995 ◽  
Vol 42 (2) ◽  
pp. 205-210 ◽  
Author(s):  
P Widłak ◽  
J Rogoliński ◽  
J Rzeszowska-Wolny
Keyword(s):  
Rat Liver ◽  
Copper Ions ◽  
Specific Interaction ◽  
Protein Composition ◽  
Matrix Proteins ◽  
Matrix Attachment Region ◽  
The Matrix ◽  
Attachment Region ◽  
Liver Nuclei ◽  
The Stability

Preincubation of rat liver nuclei with copper ions influenced the stability and protein composition of the nuclear matrices isolated by a "high salt" method. Also the specific interaction between matrix proteins and the kappa Ig matrix attachment region of DNA was affected.

scholarly journals Change in the protein composition of nuclear matrix induced by lipase treatment

1989 ◽  
Vol 5 (6) ◽  
pp. 73-77 ◽  
Author(s):  
D. Yu. Blokhin ◽  
V. A. Struchkov
Keyword(s):  
Nuclear Matrix ◽  
Protein Composition

In situ preparation of the nuclear matrix of Physarum polycephalum: ultrastructural and biochemical analysis of different matrix isolation procedures

1988 ◽  
Vol 90 (4) ◽  
pp. 621-628 ◽  
Author(s):  
W. Waitz ◽  
P. Loidl
Keyword(s):  
Nuclear Matrix ◽  
Physarum Polycephalum ◽  
Matrix Isolation ◽  
Protein Composition ◽  
Biochemical Properties ◽  
Agarose Beads ◽  
In Situ Preparation ◽  
The Matrix ◽  
Important Approach

A novel method for in situ preparation of nuclear matrix from whole plasmodia of Physarum polycephalum without isolation of nuclei is presented. Plasmodia are encapsulated in agarose beads and after solubilization of the cytoplasm the nuclear matrix is prepared. With this quick and easy technique nuclear matrix can be reproducibly prepared with perfect recovery. We compared the ultrastructural and biochemical properties of the matrix after three different matrix isolation procedures: preparation with high salt, ammonium sulphate and lithium diiodosalicylic acid. The results show that the ultrastructure and protein composition of the three types of matrix are very similar or even identical. We conclude that many of the conflicting results on nuclear matrix in the literature are due to perturbations of nuclear integrity during the isolation of nuclei. For this reason the new in situ method is an important approach in the standardization of nuclear matrix isolation.

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