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

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.

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.

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.

scholarly journals Effect of organ site on nuclear matrix protein composition

1996 ◽  
Vol 62 (1) ◽  
pp. 132-141 ◽  
Author(s):  
Tracy S. Replogle-Schwab ◽  
Robert H. Getzenberg ◽  
Terry L. Donat ◽  
Kenneth J. Pienta
Keyword(s):  
Nuclear Matrix ◽  
Matrix Protein ◽  
Protein Composition ◽  
Nuclear Matrix Protein ◽  
Organ Site

scholarly journals Progressive changes in the protein composition of the nuclear matrix during rat osteoblast differentiation.

1990 ◽  
Vol 87 (12) ◽  
pp. 4605-4609 ◽  
Author(s):  
S. I. Dworetzky ◽  
E. G. Fey ◽  
S. Penman ◽  
J. B. Lian ◽  
J. L. Stein ◽  
...  
Keyword(s):  
Nuclear Matrix ◽  
Osteoblast Differentiation ◽  
Protein Composition ◽  
Rat Osteoblast

scholarly journals Developmental association of the beta-galactoside-binding protein galectin-1 with the nuclear matrix of rat calvarial osteoblasts

1998 ◽  
Vol 111 (20) ◽  
pp. 3035-3043 ◽  
Author(s):  
J.Y. Choi ◽  
A.J. van Wijnen ◽  
F. Aslam ◽  
J.D. Leszyk ◽  
J.L. Stein ◽  
...  
Keyword(s):  
Nuclear Matrix ◽  
Osteoblast Differentiation ◽  
Matrix Protein ◽  
Binding Protein ◽  
Protein Composition ◽  
Matrix Proteins ◽  
Specific Gene ◽  
Two Dimensional Gel Electrophoresis ◽  
Nuclear Matrix Proteins ◽  
Differential Retention

The protein composition of the nuclear matrix changes significantly as the osteoblast matures from a proliferating pre-osteoblast to an osteocyte embedded in a mineralized matrix. These matrix protein are the result of developmental stage-specific gene expression during osteoblast differentiation. To isolate nuclear matrix proteins unique to the bone phenotype we analyzed nuclear matrix preparations from cultures of rat calvarial osteoblasts by high resolution two-dimensional gel electrophoresis at two different stages: proliferation (day 3) and differentiation (day 18, mineralized). We characterized one protein (14 kDa; pI 5.0), that was detectable only in the nuclear matrix of differentiated osteoblasts. By mass spectrometry and microsequencing, this protein was identified as the beta -galactoside-binding protein galectin-1. Both immunofluorescence staining of nuclear matrix preparations with the galectin-1 antibody and western blot analysis of subcellular fractions confirmed that galectin-1 is only associated with the nuclear matrix in differentiated osteoblasts as the result of differential retention. Galectin-1 protein and mRNA are present throughout osteoblast differentiation. Galectin-1 is present in the cytoplasmic and nuclear fractions in both proliferating and differentiated osteoblasts. However, its only stable binding is to the nuclear matrix of the differentiated osteoblast; but, in proliferating osteoblasts, galectin-1 is not retained in the nuclear matrix. Taken together, our results suggest that developmental association of galectin-1 with the nuclear matrix reflects differential subnuclear binding of galectin-1 during osteoblast differentiation.

The protein composition of the nuclear matrix from cells in various differentiation states: Evidence for a common set of matrix proteins

1989 ◽  
Vol 27 ◽  
pp. 225
Author(s):  
Nico Stuurnman ◽  
AlexandraM.L. Meijne ◽  
Luitzen deJong ◽  
Roel van Driel ◽  
Jos van Renswoude
Keyword(s):  
Nuclear Matrix ◽  
Protein Composition ◽  
Matrix Proteins
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