Age- and ploidy-related changes in rat liver nuclear proteins as assessed by one- and two-dimensional gel electrophoresis

1982 ◽  
Vol 60 (3) ◽  
pp. 204-214 ◽  
Author(s):  
Klaus Brasch
Keyword(s):  
Gel Electrophoresis ◽  
Minor Component ◽  
Subsequent Stage ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Nonhistone Proteins ◽  
Stage Of Development ◽  
Age Related ◽  
Associated Proteins ◽  
A Minor

Hepatocyte nuclei from young (3–5 week), mature (8–12 week), and aged (over 32 weeks) rats were isolated and characterized by flow cytometry. Nuclei were bulk separated into diploid (2C), tetraploid (4C), and octoploid (8C) enriched fractions on sucrose gradients. Total, 0.35 M NaCl soluble, and residual proteins were prepared from all nuclear stages and examined by one- and two-dimensional gel electrophoresis. Within limits of sensitivity of these techniques, the following general features emerged. (a) A majority of proteins visualized were common to and present in similar relative quantities in nuclei from all age and ploidy groups. (b) A relatively higher proportion of nonhistone proteins (NHP) were saline-soluble in 2C nuclei from young rats than at any subsequent stage of development. (c) Several age-related and to a lesser extent ploidy-related fluctuations in pattern among the NHP were evident. These reflected primarily differences in solubility rather than major quantitative changes among individual proteins. (d) Exceptions to the foregoing included a group of high molecular weight components (> 100 000), a major and a minor component between 45 000 and 50 000, and a heterogeneous group of proteins in 2C nuclei from very young animals. There were no obvious differences among the histones, although these proteins were not examined in detail. The complex pattern of changes observed are discussed in terms of known aspects of hepatocyte differentiation and are related to possible changes in nucleoplasmic, nuclear matrix and Hn-RNP associated proteins.

scholarly journals A search for differential polypeptide synthesis throughout the cell cycle of HeLa cells.

1980 ◽  
Vol 84 (3) ◽  
pp. 795-802 ◽  
Author(s):  
R Bravo ◽  
J E Celis
Keyword(s):  
Cell Cycle ◽  
Hela Cells ◽  
Gel Electrophoresis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Nonhistone Proteins ◽  
Polypeptide Synthesis ◽  
Acidic Proteins ◽  
Filament Protein

The polypeptides synthesized during the cell cycle of HeLa cells were analyzed by means of two-dimensional gel electrophoresis followed by fluorography under conditions in which the position of 700 polypeptides (acidic and basic) could be reproducibly assessed. Mitotic cells obtained by mechanical detachment and synchronized cells in other stages of the cell cycle were labeled with [35S]methionine for 30-min pulses or for long terms starting at the beginning of each phase. Visual comparison of the polypeptide maps obtained in the different stages of the cell cycle showed that these were strikingly similar, and there was no indication that the synthesis of any of the detected polypeptides was confined to only one of the cell cycle phases. Quantitation of 99 abundant polypeptides (acidic and basic) in pulse-labeled and long-term labeled cells revealed that the relative amount (i.e., the rate of synthesis) of most polypeptides, including total actin, alpha-actinin, 6 abundant basic nonhistone proteins, and 13 major acidic proteins present in Triton cytoskeletons, remains constant throughout the cell cycle. Among the few variable polypeptides (markers), we have identified alpha- and beta-tubulin (increase in M), the subunit of the 100-A filament protein "fibroblast type" (decreases in M), and a 36,000 mol wt acidic cytoarchitectural protein that increases in S. A few other unidentified polypeptides have also been found to vary in M and in M and G2, but no marker was found in G1.

scholarly journals RNA-seq and two-dimensional gel electrophoresis revealed genomic and proteomic signatures during bacteriophage-host interactions in Pseudomonas aeruginosa

2019 ◽  
Author(s):  
Yinling Tan ◽  
Qiu Zhong ◽  
Xia Zhao ◽  
Canhuang Chen ◽  
Yang Li ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Gel Electrophoresis ◽  
Phage Infection ◽  
Two Dimensional ◽  
Rna Seq ◽  
Two Dimensional Gel Electrophoresis ◽  
Host Interactions ◽  
Associated Proteins ◽  
Infection Processes ◽  
Lysogenic Phage

Abstract Background Understanding the biological nature of bacteriophage is important in exploring the therapeutic and biotechnological potentials of bacteriophages. However, available information is limited to the infection processes on either model phages infecting Escherichia coli or lytic phages against pathogens. The interplay between lysogenic phage and its host was rarely studied. Results We investigated the interactions between Pseudomonas aeruginosa and a lysogenic bacteriophage PaP3 through RNA-seq and two-dimensional gel electrophoresis (2D-GE). Compared to the uninfected host, a total of 2,891 (51.3%) differentially expressed genes (DGEs) were identified, most of which were repressed by phages, including the changes in metabolic-related and virulence-associated genes. The RT-qPCR results showed consistent directional changes compared with the RNA-seq results. According to 2D-GE, phage structure proteins were detected after phage infection. The host proteins, such as flagella hook-associated proteins, disappeared gradually after phage infection and may be shut off by phage. Conclusions All these indicate that although lysogenic phages do not immediately lyse the host, they play a significant regulatory role in the expression of host genes. Our findings provide an expanded view of the lysogenic phage infection processes and may offer potential targets for therapeutic intervention against P. aeruginosa infections.

Two-dimensional gel electrophoresis can be used to examine chromatographic effluent fractions from displacement column chromatography.

1982 ◽  
Vol 28 (4) ◽  
pp. 998-999 ◽  
Author(s):  
A R Torres ◽  
G G Krueger ◽  
E A Peterson
Keyword(s):  
Human Serum ◽  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Gel Method ◽  
Minor Protein ◽  
Protein Components ◽  
A Minor

Abstract We show how two-dimensional gel electrophoresis can be used to monitor protein components in effluent fractions from a displacement column. A minor protein in human serum, of interest in studies on psoriasis and highly enriched by using carboxymethyldextrans as displacers on DEAE-Sephacel, was easily detected in the effluent fractions with the two-dimensional gel method because its concentration was sufficiently high and there was no interference by the carboxymethyldextrans or salt.

Diversity of Glycoprotein Deficiencies in Glanzmann’s Thrombasthenia

1985 ◽  
Vol 54 (03) ◽  
pp. 626-629 ◽  
Author(s):  
M Meyer ◽  
F H Herrmann
Keyword(s):  
High Resolution ◽  
Gel Electrophoresis ◽  
Type Ii ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Structural Variant ◽  
Glanzmann’S Thrombasthenia ◽  
Crossed Immunoelectrophoresis ◽  
Glanzmann's Thrombasthenia ◽  
Platelet Proteins

SummaryThe platelet proteins of 9 thrombasthenic patients from 7 families were analysed by high resolution two-dimensional gel electrophoresis (HR-2DE) and crossed immunoelectrophoresis (CIE). In 7 patients both glycoproteins (GPs) IIb and Ilia were absent or reduced to roughly the same extent. In two related patients only a trace of GP Ilb-IIIa complex was detected in CIE, but HR-2DE revealed a glycopeptide in the position of GP Ilia in an amount comparable to type II thrombasthenia. This GP Ilia-like component was neither recognized normally by anti-GP Ilb-IIIa antibodies nor labeled by surface iodination. In unreduced-reduced two-dimensional gel electrophoresis two components were observed in the region of GP Ilia. The assumption of a structural variant of GP Ilia in the two related patients is discussed.

Sign in / Sign up

Export Citation Format

Share Document