Changes in patterns of protein synthesis in axolotl oocytes during progesterone-induced maturation

Development ◽  
1986 ◽  
Vol 92 (1) ◽  
pp. 103-113
Author(s):  
Jean Gautier ◽  
Renée Tencer
Keyword(s):  
Protein Synthesis ◽  
Protein Phosphorylation ◽  
Cholera Toxin ◽  
Oocyte Maturation ◽  
Gel Electrophoresis ◽  
De Novo ◽  
Protein Pattern ◽  
Ambystoma Mexicanum ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis

Patterns of protein phosphorylation and synthesis during axolotl (Ambystoma mexicanum) oocyte maturation were studied by incorporation of [32P]orthophosphate and [35S]methionine into polypeptides, followed by two-dimensional gel electrophoresis. Various alterations were observed after progesterone treatment: de novo appearance of [35S]methionine-labelled polypeptides, a quantitative increase in previously synthesized proteins and a quantitative decrease in or disappearance of other previously synthesized proteins. Changes in 32P- and 35S-labelling were observed very early during maturation. Neither prior oocyte enucleation nor α-amanitin treatment had a significant effect on these changes. Stimulation with MPF provided the same final protein pattern as PG treatment. However, cholera toxin inhibited all the changes seen during maturation. Comparisons between the patterns of [35S]methionine- and [32P]phosphatelabelling provide further information on the biochemical events that take place during oocyte maturation.

Two-dimensional gel electrophoresis of serum specimens from a normal population.

1982 ◽  
Vol 28 (4) ◽  
pp. 890-899 ◽  
Author(s):  
R P Tracy ◽  
R M Currie ◽  
D S Young
Keyword(s):  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Protein Pattern ◽  
Normal Population ◽  
Small Samples ◽  
Two Dimensional ◽  
Silver Stain ◽  
Two Dimensional Gel Electrophoresis ◽  
Finger Stick ◽  
Effects Of Aging

Abstract We examined sera from a normal population by two-dimensional gel electrophoresis, to establish the normal pattern of serum proteins and to investigate genetic polymorphisms. With such information in hand, specimens from patients with certain diseases may be readily evaluated. Towards this goal, we optimized the ISO-DALT system (Proc. Natl, Acad, Sci, USA 74: 5421--5425, 1977) for routine phenotyping of alpha 1-antitrypsin, haptoglobin, GC-globulin, alpha 2-HS-glycoprotein, and transferrin, as well as a previously unknown polymorphic protein. We examined the effects of aging the specimens for 2 h at room temperature (no changes) or at -20 degrees C for several months (small changes), as well as serum/plasma differences and the effect of protease inhibitors. Silver-stain methods were modified to allow simultaneous staining of 10 gels, with reasonably good reproducibility of stain intensity. We quantitated silver-stained gels by densitometry of photographic transparencies. Very small samples suffice with this stain (0.5 microL of serum or plasma), allowing the use of "finger-stick" methods instead of venipuncture, yet the patterns are better resolved and easier to read than those for 10-microL specimens processed on gels stained with Coomassie Blue. Our techniques for rapidly removing albumin and IgG allow the investigator to examine areas on the gel that ordinarily are obscured. The region of haptoglobin has been examined by using serum from an ahaptoglobinemic donor. Finally, we present an expanded "normal" map illustrating the composite protein pattern.

Cytoskeletal proteins from human skin fibroblasts, peripheral blood leukocytes, and a lymphoblastoid cell line compared by two-dimensional gel electrophoresis.

1982 ◽  
Vol 28 (4) ◽  
pp. 955-961 ◽  
Author(s):  
C S Giometti ◽  
K E Willard ◽  
N L Anderson
Keyword(s):  
Gel Electrophoresis ◽  
Lymphoblastoid Cell Line ◽  
Protein Pattern ◽  
Cytoskeletal Proteins ◽  
Cytoskeletal Protein ◽  
Two Dimensional ◽  
Human Skin Fibroblasts ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Two Dimensional Gel Electrophoresis

Abstract Differences in proteins between cells grown as suspension cultures and those grown as attached cultures were studied by comparing the proteins of detergent-resistant cytoskeletons prepared from peripheral blood leukocytes and a lymphoblastoid cell line (GM607) (both grown as suspension cultures) and those of human skin fibroblasts (grown as attached cultures) by two-dimensional gel electrophoresis. The major cytoskeletal proteins of the leukocytes were also present in the protein pattern of GM607 cytoskeletons. In contrast, the fibroblast cytoskeletal protein pattern contained four groups of proteins that differed from the patterns of the leukocytes and GM607. Three groups (Cytoskf:8--10, :14--16, and :17--18) showed qualitative differences, and the fourth group (Cytoskf:11 and :13) showed quantitative differences. In addition, surface labeling of GM607 and human fibroblasts with 125I demonstrated that substantial amounts of vimentin and actin are exposed at the surface of the attached fibroblasts, but there is little evidence of similar exposure at the surface of the suspension-grown GM607. Cytoskf:11 and :13 in fibroblast preparations were also labeled with the 125I. These results demonstrate some differences in cytoskeletal protein composition between different types of cells could be related to their ability or lack of ability to grow as attached cells in tissue culture.

scholarly journals Multiple forms of tubulin in the cytoskeletal and flagellar microtubules of polytomella

1981 ◽  
Vol 91 (2) ◽  
pp. 352-360 ◽  
Author(s):  
TW McKeithan ◽  
JL Rosenbaum
Keyword(s):  
Protein Synthesis ◽  
Gel Electrophoresis ◽  
Cold Temperature ◽  
Two Dimensional ◽  
Multiple Forms ◽  
Cytoplasmic Fraction ◽  
Two Dimensional Gel Electrophoresis ◽  
Brain Tubulin ◽  
Β Tubulin

The alga polytomella contains several organelles composed of microtubules, including four flagella and hundreds of cytoskeletal microtubules. Brown and co-workers have shown (1976. J. Cell Biol. 69:6-125; 1978, Exp. Cell Res. 117: 313-324) that the flagella could be removed and the cytoskeletans dissociated, and that both structures could partially regenerate in the absence of protein synthesis. Because of this, and because both the flagella and the cytoskeletons can be isolated intact, this organism is particularly suitable for studying tubulin heterogeneity and the incorporation of specific tubulins into different microtubule-containing organelles in the same cell. In order to define the different species of tubulin in polytonella cytoplasm, a (35)S- labeled cytoplasmic fraction was subjected to two cycles of assembly and disassembly in the presence of unlabeled brain tubulin. Comparison of the labeled polytomella cytoplasmic tubulin obtained by this procedure with the tubulin of isolated polytomella flagella by two-dimensional gel electrophoresis showed that, whereas the β-tubulin from both cytoplasmic and flagellar tubulin samples comigrated, the two α-tubulins had distinctly different isoelectic points. As a second method of isolating tubulin from the cytoplasm, cells were gently lysed with detergent and intact cytoskeletons obtained. When these cytoskeletons were exposed to cold temperature, the proteins that were released were found to be highly enriched in tubulin; this tubulin, by itself, could be assembled into microtubules in vitro. The predominant α-tubulin of this in vitro- assembled cytoskeletal tubulin corresponded to the major cytoplasmic α-tubulin obtained by coassembly of labeled polytomella cytoplasmic extract with brain tubulin and was quite distinct from the α-tubulin of purified flagella. These results clearly show that two different microtubule-containing organelles from the same cell are composed of distinct tubulins.

Effects of selenium deficiency on the rat myocardial protein pattern - investigation by two-dimensional gel electrophoresis

2000 ◽  
Vol 95 (3) ◽  
pp. 199-207 ◽  
Author(s):  
Vera Regitz-Zagrosek ◽  
Antonius Kyriakopoulos ◽  
Peter Jungblut ◽  
Dietrich Behne ◽  
Klaus-Peter Plei�ner ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Protein Pattern ◽  
Selenium Deficiency ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis

Protein Synthesis During Oxygen Deprivation in Cotton

1996 ◽  
Vol 23 (3) ◽  
pp. 341 ◽  
Author(s):  
AA Millar ◽  
ES Dennis
Keyword(s):  
Molecular Weight ◽  
Protein Synthesis ◽  
Gel Electrophoresis ◽  
Major Part ◽  
Oxygen Deprivation ◽  
Two Dimensional ◽  
Root Tips ◽  
Two Dimensional Gel Electrophoresis ◽  
Molecular Masses ◽  
Adh Activity

The alteration of protein synthesis induced by oxygen deprivation has been examined in the root tips of cotton (Gossypium hirsutum cv. Siokra), a plant that is intolerant to anoxia. Using [35S]methionine labelling and two-dimensional gel electrophoresis it was demonstrated that 14 major polypeptides are being selectively synthesised during oxygen deprivation. These polypeptides have been designated the cotton anaerobic polypeptides (ANPs), and have estimated molecular masses that correspond to molecular masses of ANPs from other plant species. However, compared to maize, several of the major molecular weight classes are absent, suggesting that the response to oxygen deprivation in cotton is simpler than that of maize. Alcohol dehydrogenase (ADH) activity is induced by oxygen deprivation. Using western analysis we have determined that this increase in activity is correlated with the accumulation of the ADH polypeptide and that three of the major cotton ANPs are ADH, including the most intensely labelled ANP, demonstrating that the synthesis of ADH constitutes a major part of the response in cotton.

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