scholarly journals The subunit structure of the arom multienzyme complex of Neurospora crassa. Evidence from peptide ‘maps’ for the identity of the subunits

1978 ◽  
Vol 169 (2) ◽  
pp. 441-444 ◽  
Author(s):  
J Lumsden ◽  
J R Coggins
Keyword(s):  
Neurospora Crassa ◽  
Gel Electrophoresis ◽  
Peptide Mapping ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Subunit Structure ◽  
Multienzyme Complex ◽  
Peptide Maps

Evidence was obtained, from polyacrylamide-gel electrophoresis in the presence of urea and from peptide ‘mapping’ of specifically labelled cysteine-and methionine-containing peptides, that the two subunits of the arom multienzyme complex of Neurospora crassa are chemically very similar and possibly identical.

scholarly journals Tubulin heterogeneity in the trypanosome Crithidia fasciculata.

1984 ◽  
Vol 4 (4) ◽  
pp. 779-790 ◽  
Author(s):  
D G Russell ◽  
D Miller ◽  
K Gull
Keyword(s):  
Gel Electrophoresis ◽  
Peptide Mapping ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
In Vitro Translation ◽  
Two Dimensional ◽  
Post Translational Modification ◽  
Interphase Cell ◽  
Crithidia Fasciculata ◽  
Alpha Tubulin

The interphase cell of Crithidia fasciculata has three discrete tubulin populations: the subpellicular microtubules, the axonemal microtubules, and the nonpolymerized cytoplasmic pool protein. These three tubulin populations were independently and selectively purified, yielding, in each case, microtubule protein capable of self-assembly. All three preparations polymerized to form ribbons and sheets rather than the more usual microtubular structures. Analyses of the tubulin by two-dimensional polyacrylamide gel electrophoresis, isoelectric focusing, and peptide mapping indicated that the beta-tubulin complex remained constant regardless of source but that some heterogeneity was present in the alpha subunit. Cytoplasmic pool alpha tubulins (alpha 1/alpha 2) were the only alpha isotypes in the cytoplasm and also formed most of the alpha tubulin species in the pellicular fraction. Flagellar alpha tubulin (alpha 3) was the sole alpha isotype in the flagella; it appeared in small amounts in the pellicular fraction but was completely absent from the cytoplasm. In vitro translation products from polyadenylated RNA from C. fasciculata were also examined by two-dimensional polyacrylamide gel electrophoresis and possessed a protein corresponding to alpha 1/alpha 2 tubulin but lacked any alpha 3 tubulin. The alpha 3 polypeptide arose from a post-translational modification of a precursor polypeptide not identifiable by two-dimensional polyacrylamide gel electrophoresis as alpha 3. Peptide mapping data indicated that cytoplasmic alpha tubulin is the most likely precursor. These results demonstrate alpha-tubulin heterogeneity in this organism and also how close the relationship between flagellar and cytoskeletal tubulins can be among lower eucaryotes.

scholarly journals Purification and subunit structure of legumin of Vicia faba L. (broad bean)

1974 ◽  
Vol 141 (2) ◽  
pp. 413-418 ◽  
Author(s):  
David J. Wright ◽  
Donald Boulter
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Broad Bean ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Exchange Chromatography ◽  
Polyacrylamide Gel ◽  
Subunit Structure ◽  
Isoelectric Precipitation ◽  
Molecular Weights

Zonal isoelectric precipitation was shown to be an effective method for the preparation of legumin which was homogeneous as judged by ultracentrifugation and polyacrylamide-gel electrophoresis. The subunit structure of legumin was investigated by preparative sodium dodecyl sulphate–polyacrylamide-gel electrophoresis and ion-exchange chromatography in urea. Five distinct subunits, of which two were acidic (α) and had a molecular weight of 37000, and three were basic (β) with molecular weights of 20100, 20900 and 23800, were identified. The α and β subunits were present in equimolar amounts in the legumin molecule and, in view of this and molecular-weight considerations, an α6β6 subunit model was proposed for legumin.

scholarly journals A set of surface proteins common to the circulating human platelet and lymphocyte

1974 ◽  
Vol 141 (3) ◽  
pp. 909-911 ◽  
Author(s):  
M. J. A. Tanner ◽  
D. H. Boxer ◽  
Jane Cumming ◽  
J. Verrier-Jones
Keyword(s):  
Gel Electrophoresis ◽  
Human Platelet ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Surface Proteins ◽  
Major Type ◽  
Cell Type ◽  
Peptide Maps

The surface proteins of the circulating human platelet and lymphocyte were labelled by using the lactoperoxidase iodination method. Polyacrylamide-gel electrophoresis showed that four corresponding labelled proteins are found on the surface of each cell type. The most intensely labelled protein contains little or no carbohydrate, but the remaining labelled proteins are all glycoproteins. The major labelled band from each cell was isolated and comparative peptide ‘maps’ showed that the two proteins are closely similar. The surface proteins of the lymphocyte and platelet are distinct from those on the erythrocyte, the remaining major type of circulating cell.

Tubulin heterogeneity in the trypanosome Crithidia fasciculata

1984 ◽  
Vol 4 (4) ◽  
pp. 779-790
Author(s):  
D G Russell ◽  
D Miller ◽  
K Gull
Keyword(s):  
Gel Electrophoresis ◽  
Peptide Mapping ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
In Vitro Translation ◽  
Two Dimensional ◽  
Post Translational Modification ◽  
Interphase Cell ◽  
Crithidia Fasciculata ◽  
Alpha Tubulin

The interphase cell of Crithidia fasciculata has three discrete tubulin populations: the subpellicular microtubules, the axonemal microtubules, and the nonpolymerized cytoplasmic pool protein. These three tubulin populations were independently and selectively purified, yielding, in each case, microtubule protein capable of self-assembly. All three preparations polymerized to form ribbons and sheets rather than the more usual microtubular structures. Analyses of the tubulin by two-dimensional polyacrylamide gel electrophoresis, isoelectric focusing, and peptide mapping indicated that the beta-tubulin complex remained constant regardless of source but that some heterogeneity was present in the alpha subunit. Cytoplasmic pool alpha tubulins (alpha 1/alpha 2) were the only alpha isotypes in the cytoplasm and also formed most of the alpha tubulin species in the pellicular fraction. Flagellar alpha tubulin (alpha 3) was the sole alpha isotype in the flagella; it appeared in small amounts in the pellicular fraction but was completely absent from the cytoplasm. In vitro translation products from polyadenylated RNA from C. fasciculata were also examined by two-dimensional polyacrylamide gel electrophoresis and possessed a protein corresponding to alpha 1/alpha 2 tubulin but lacked any alpha 3 tubulin. The alpha 3 polypeptide arose from a post-translational modification of a precursor polypeptide not identifiable by two-dimensional polyacrylamide gel electrophoresis as alpha 3. Peptide mapping data indicated that cytoplasmic alpha tubulin is the most likely precursor. These results demonstrate alpha-tubulin heterogeneity in this organism and also how close the relationship between flagellar and cytoskeletal tubulins can be among lower eucaryotes.

scholarly journals Dog and human acid β-d-galactosidases are structurally similar

1983 ◽  
Vol 213 (2) ◽  
pp. 473-478 ◽  
Author(s):  
J J Hubert ◽  
J S O′Brien
Keyword(s):  
Gel Electrophoresis ◽  
Human Liver ◽  
Peptide Mapping ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Major Band ◽  
Chymotryptic Peptide ◽  
Human Acid ◽  
Dog Liver ◽  
Peptide Maps

The purification of dog liver acid β-galactosidase is described. The dog enzyme migrated as a single major band on polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate, with a molecular weight of 60000. Antiserum raised against purified human liver acid β-galactosidase cross-reacted with β-galactosidase from dog liver, but not with those from cat liver or Escherichia coli. Tryptic peptide maps of the dog and human acid β-galactosidases indicate that 21 of the 24 peptides observed were homologous; a similar result was obtained after chymotryptic peptide mapping. We conclude that dog and human acid β-galactosidases are structurally similar, and that canine GM1 gangliosidosis (acid β-galactosidase deficiency) is an excellent model for the same disease in man.

γ’-Dimer Antibody and its Specificity Against Fibrinogen, FR-a and Carboxymethylated chains from Fibrinogen and Cross-linked Fibrin

1975 ◽  
Author(s):  
N. T. Clausen ◽  
J. Gormsen
Keyword(s):  
Ionic Strength ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Acetate Buffer ◽  
Subunit Structure ◽  
Weak Reaction ◽  
Subcutaneous Injections ◽  
L 96 ◽  
Heterologous Antibody

γ’-dimers (M: 80.000) were obtained from fully cross-linked fibrin (Kabi, grade L, 96% clottable) by digestion with porcine plasmin (0.5 CTA units/mg protein) for 18 h., carboxymethylation and subsequent isolation on Whatman CM-52 with a combined pH (4.8 → 5.5) and ionic strength (0.05 → 0.25 M) gradient in Na-acetate buffer with 8 M urea*).The γ’-dimers were recycled once and exhibited purity on SDS polyacrylamide gel electrophoresis. These γ’-dimers showed weak reaction in immunoprecipitation assays with anti-fibrinogen and anti-D but not with anti-E rabbit immunserum.Heterologous antibody was obtained from rabbit after subcutaneous injections of γ’-dimer antigen and bleeding after 4 months, 14 days after last injection.This crude antibody reacted in immunodiffusion and monorocket immunoelectrophoresis with γ’-dimers and weakly with fibrinogen and fragment D but not with fragment E (DEAE purified). After absorption with fibrinogen and fractional desalting with (NH4)2SO4 (25% and 50%) there was no reaction with fibrinogen, D or E fragments, but still with γ’-dimers.Testing against isolated chains (SDS-pure) from fibrinogen revealed Aα cross-reaction but only reaction with γ and γ’-dimer chains remained after proper absorption. Possible differentiation between these two subunit structure is reported.* Murano, G. et al. FEBS-letters 14, 37–41, 1971.

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