Identification of concanavalin A-binding proteins after sodium dodecyl sulfate-gel electrophoresis and protein blotting

1982 ◽  
Vol 123 (1) ◽  
pp. 143-146 ◽  
Author(s):  
Richard Hawkes
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Concanavalin A ◽  
Gel Electrophoresis ◽  
Binding Proteins ◽  
Sodium Dodecyl ◽  
Protein Blotting ◽  
Dodecyl Sulfate

Identification of liver plasma membrane glycoproteins which bind to 125I-Labelled concanavalin A following electrophoresis in sodium dodecyl sulfate

1976 ◽  
Vol 54 (5) ◽  
pp. 477-480 ◽  
Author(s):  
James W. Gurd ◽  
W. Howard Evans
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Concanavalin A ◽  
Gel Electrophoresis ◽  
Plasma Membranes ◽  
Sodium Dodecyl ◽  
Membrane Composition ◽  
Membrane Glycoproteins ◽  
Con A ◽  
General Application ◽  
Dodecyl Sulfate

Following electrophoresis of ovalbumin in sodium dodecyl sulfate (SDS) this glycoprotein bound 125I-labelled concanavalin A (Con A). The reaction was specific and proportional to the amount of glycoprotein present on the gel. This technique was used to study the Con-A-binding glycoproteins of liver cell surfaces. Mouse liver plasma membranes were purified and subfractionated to yield two fractions corresponding to the bile canalicular surface and the surface between adjacent hepatocytes (Evans, W. H. (1970) Biochem. J. 116, 833–842). Both fractions bound 125I-labelled Con A, the former binding two to three times more lectin than the latter. Following SDS gel electrophoresis individual membrane glycoproteins reacted with 125I-labelled Con A. Both membrane subfractions yielded qualitatively similar Con A binding profiles, seven binding proteins being present in each. The results are consistent with a generally uniform distribution of glycoproteins over the hepatocyte surface. The reaction of lectins with glycoproteins following SDS gel electrophoresis should find general application in the study of membrane composition.

DNA binding proteins from Tetrahymena thermophila

1982 ◽  
Vol 60 (3) ◽  
pp. 398-407
Author(s):  
Nora N. G. Tsao ◽  
Ronald E. Pearlman
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Dna Polymerase ◽  
Gel Electrophoresis ◽  
Binding Proteins ◽  
Tetrahymena Thermophila ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Dodecyl Sulfate

We have used DNA-cellulose chromatography to isolate single-strand binding proteins from Tetrahymena thermophila. Three major proteins which bind to denatured DNA-cellulose were obtained. The predominant protein has a molecular weight of 20 000 in sodium dodecyl sulfate – polyacrylamide gel electrophoresis and possesses many of the properties of the helix destabilizing proteins isolated from prokaryotic and eukaryotic sources. The protein facilitates denaturation of the synthetic copolymer poly[d(A-T)∙d(A-T)], depressing the melting temperature by nearly 40 °C. It also permits the renaturation of poly[d(A-T)∙d(A-T)] in high salt concentration. Two other binding proteins have molecular weights of 25 000 and 23 000 in sodium dodecyl sulfate – polyacrylamide gel electrophoresis. The protein with a molecular weight of 25 000 is probably the "M protein" previously isolated from Tetrahymena thermophila which has been shown to stimulate Tetrahymena DNA polymerase. These two proteins failed to show helix destabilizing, DNA dependent ATPase, or deoxyribonuclease activities. These three proteins are abundant in the cell with approximately 1.0 × 106 to 10.0 × 106 molecules of each protein monomer per cell. One molecule of each protein monomer binds to 7 to 10 nucleotides as detected by a nitrocellulose filter binding assay. Peptide mapping of the three proteins suggests that they are all distinct. We have also found that the binding proteins can interact with Tetrahymena DNA polymerase and some other proteins to form an enzyme complex, a putative replication complex.

Effect of thyroid hormone on carbohydrate content of Na+-K+-adenosine triphosphatase

1984 ◽  
Vol 247 (3) ◽  
pp. C282-C287 ◽  
Author(s):  
C. S. Lo ◽  
L. E. Klein ◽  
T. N. Lo
Keyword(s):  
Thyroid Hormone ◽  
Sodium Dodecyl Sulfate ◽  
Rate Constant ◽  
Gel Electrophoresis ◽  
Adenosine Triphosphatase ◽  
Sodium Dodecyl ◽  
Carbohydrate Content ◽  
Beta Subunits ◽  
Reverse T3 ◽  
Dodecyl Sulfate

The effect of L-3,5,3'-triiodothyronine (T3) (50 micrograms/100 body wt) on the incorporation of labeled glucosamine and fucose into the subunits of Na+-K+-ATPase was examined by gel electrophoresis in sodium dodecyl sulfate. T3 augmented the incorporation of glucosamine into the alpha- and beta-subunits by 51 and 58%, respectively, in the 22-h chase experiments. Similarly T3 augmented the incorporation of fucose into the alpha- and beta-subunits by 58 and 43%, respectively. Reverse T3 did not alter the incorporation of labeled fucose in either subunit. The effect of T3 on the rate constant of degradation of renal cortical Na+-K+-ATPase was assessed. The rate constant of degradation (Kd) of the [3H]fucose labeled alpha- and beta-subunits for the hypothyroid rats were both 0.20, and for T3-treated rats, the Kd of the alpha- and beta-subunits were 0.23 and 0.18, respectively, suggesting that T3 enhanced fucose incorporation into the subunits of Na+-K+-ATPase rather than retarding the degradation of this enzyme.

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