scholarly journals Preparation, proteolysis and reversible oxidation of highly purified Azotobacter vinelandii polynucleotide phosphorylase

1970 ◽  
Vol 120 (4) ◽  
pp. 753-761 ◽  
Author(s):  
A. T. Gajda ◽  
G. Zaror de Behrens ◽  
P. S. Fitt
Keyword(s):  
Electrophoretic Mobility ◽  
Good Yield ◽  
Azotobacter Vinelandii ◽  
New Method ◽  
Reduced Form ◽  
Polyacrylamide Gels ◽  
Polynucleotide Phosphorylase ◽  
Oligonucleotide Primers ◽  
Aerial Oxidation

1. A new method has been developed for the preparation in good yield of highly purified Azotobacter vinelandii polynucleotide phosphorylase in its reduced form. 2. Aging or digestion with trypsin causes the enzyme to develop a primer requirement that is not eliminated by β-mercaptoethanol. 3. The development of a primer requirement is accompanied by marked changes of the electrophoretic mobility of the enzyme in polyacrylamide gels. 4. The enzyme is inactivated by aerial oxidation or thiol-specific reagents. The lost activity is restored by β-mercaptoethanol, but not by oligonucleotide primers.

scholarly journals A study by polyacrylamide-gel electrophoresis of the effect of proteolysis on Micrococcus lysodeikticus polynucleotide phosphorylase

1968 ◽  
Vol 110 (3) ◽  
pp. 475-479 ◽  
Author(s):  
P. S. Fitt ◽  
E. A. Fitt ◽  
H. Wille
Keyword(s):  
Electrophoretic Mobility ◽  
Degradation Product ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Progressive Increase ◽  
Active Species ◽  
Polyacrylamide Gel ◽  
Polyacrylamide Gels ◽  
Polynucleotide Phosphorylase ◽  
Micrococcus Lysodeikticus

1. Trypsin digestion of Micrococcus lysodeikticus polynucleotide phosphorylase (nucleoside diphosphate–polynucleotide nucleotidyltransferase) causes a progressive increase in electrophoretic mobility in polyacrylamide gels of the single active degradation product. 2. A marked increase in primer requirement for CDP polymerization occurs before a more mobile product is formed. 3. α-Chymotrypsin digestion yields a product that separates into several active species on polyacrylamide-gel electrophoretograms. 4. No separation of ADP-and CDP-polymerization activities occurs during electrophoresis after either trypsin or α-chymotrypsin treatment.

A comparative study of the insoluble storage proteins and the lectins of seeds of the Euphorbiaceae

1984 ◽  
Vol 62 (8) ◽  
pp. 1671-1677 ◽  
Author(s):  
Louise Lalonde ◽  
David W. Fountain ◽  
Allison Kermode ◽  
Francis B. Ouellette ◽  
Kelly Scott ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Castor Bean ◽  
Storage Proteins ◽  
Reduced Form ◽  
Polyacrylamide Gels ◽  
Salt Solutions ◽  
Sodium Dodecylsulphate ◽  
Molecular Weight Range ◽  
Jatropha Gossypifolia ◽  
Major Storage Protein

The major storage protein within seeds of the Euphorbiaceae is the 11S crystalloid, which is only completely soluble in buffer or salt solutions if sodium dodecylsulphate or urea is present. Prior to this study, only the storage proteins of the castor bean had been characterized. The nonreduced crystalloid protein complex in all species tested has a molecular weight of 50 000 – 55 000, and in reduced form the proteins migrate on polyacrylamide gels as two distinct groups of polypeptides, one in the molecular weight range 20 000 – 25 000 and the other in the 29 000 – 35 000 range. In this respect the proteins have the general characteristics of those of castor bean, but only the proteins of Jatropha gossypifolia show striking similarities. Within any one genus, the storage proteins appear to be more or less identical (e.g., Manihot spp.) or show distinct differences (e.g., Euphorbia spp.). The soluble lectin proteins of J. gossypifolia have very similar haemagglutination properties to those of castor bean lectins, and the glycoproteins of both species separate similarly on polyacrylamide gels. Few other species contain glycoproteins or lectins that can cause agglutination.

Glycophorin expression in murine erythroleukaemia cells

1989 ◽  
Vol 92 (2) ◽  
pp. 163-171
Author(s):  
J.B. Ulmer ◽  
E.D. Dolci ◽  
G.E. Palade
Keyword(s):  
Cell Line ◽  
Electrophoretic Mobility ◽  
Normal Mouse ◽  
Fatty Acyl ◽  
Dimethyl Sulphoxide ◽  
Partial Characterization ◽  
Polyacrylamide Gels ◽  
Putative Precursor

We have identified mature and putative precursor forms of glycophorins expressed in a virus-transformed murine erythroleukaemia (MEL) cell line and compared them with their normal erythroblast counterparts. The following differences were found: (1) the two major MEL cell glycophorins (apparent Mr values 29–30 and 43(x10(3] have greater mobility on polyacrylamide gels than their normal gp-3 and gp-2 counterparts, due at least in part to differences in their oligosaccharide sidechains; (2) MEL cell gp-3 consists of two discrete proteins; and (3) there are more potential glycophorin precursors in MEL cells than in normal mouse erythroblasts. Four proteins, with apparent Mr values of 21, 23, 26 and 27(x10(3], have tentatively been identified as glycophorin precursors, based on the following findings: (1) they are immunologically related to the glycophorins; and (2) their synthesis was induced by dimethyl sulphoxide coincidentally with that of gp-3 and gp-2. They do not appear to be glycoproteins, as evidenced by their lack of incorporation of [3H]galactose, [3H]glucosamine or [3H]mannose. In contrast, gp-3 and gp-2 incorporated [3H]galactose and [3H]glucosamine but not [3H]mannose. Partial characterization of the glycan moieties of MEL cell glycophorins indicates that they consist mostly of tri- and tetrasaccharides, with no indication of any N-linked chains. Hence, the glycans of MEL cell glycophorins are mostly (if not all) O-linked. Furthermore, treatment with N-glycanase did not change their electrophoretic mobility on polyacrylamide gels. MEL cell glycophorins were also shown to be modified by phosphoryl and fatty acyl groups.

Comparative Enzymatic Degradation of H1 Subfractions from Syrian Hamster Tissues

1986 ◽  
Vol 41 (7-8) ◽  
pp. 776-780
Author(s):  
Elżbieta Hrabec ◽  
Anna Płucienniczak ◽  
Henryk Panusz
Keyword(s):  
Electrophoretic Mobility ◽  
Syrian Hamster ◽  
Cleavage Site ◽  
Enzymatic Degradation ◽  
Histone H1 ◽  
Polyacrylamide Gels ◽  
H1 Histone ◽  
Bothrops Marajoensis ◽  
Bothrops Moojeni

Abstract An additional hydrolysis site recognized by thrombin on histone H1 molecules was found. Snakes venom proteases from Agkistrodon rhodostoma, Bothrops marajoensis and Bothrops moojeni were further used for the analysis of H1 histones. The presence of the main cleavage site on H1 histone molecules has been established. This site is localized on main N-terminal thrombin peptide. The main venom protease peptides obtained from different H1 subfractions preserve differences of electrophoretic mobility in acid-urea polyacrylamide gels typical for the initial H1 subfractions.

A new method for the detection of enolase activity on polyacrylamide gels

1979 ◽  
Vol 98 (1) ◽  
pp. 226-230 ◽  
Author(s):  
H.K. Sharma ◽  
Morton Rothstein
Keyword(s):  
New Method ◽  
Polyacrylamide Gels

Concanavalin A-horseradish peroxidase bridge staining of alpha-1 glycoproteins separated by isoelectric focusing on polyacrylamide gel.

1976 ◽  
Vol 24 (8) ◽  
pp. 908-914 ◽  
Author(s):  
R C Allen ◽  
S S Spicer ◽  
D Zehr
Keyword(s):  
Horseradish Peroxidase ◽  
Isoelectric Focusing ◽  
Concanavalin A ◽  
Periodic Acid ◽  
Polyacrylamide Gel ◽  
New Method ◽  
Polyacrylamide Gels ◽  
Periodic Acid Schiff ◽  
Coomassie Blue

The Coomassie Blue protein stain and the periodic acid-Schiff stain for glycoproteins are compared to a new method of staining glycoproteins resolved electrophoretically. The method utilizes a Concanavalin A-horseradish peroxidase sequence to visualize selectively glycoproteins with terminal or internal mannose or terminal N-acetylglucosamine. The method applied to characterization of M and Z allele products of alpha-l-antitrypsins separated by isoelectric focusing of polyacrylamide gels slabs have revealed differences in carbohydrate content of various components that were previously undetected.

A new method for identification of proteins separated in Polyacrylamide gels

1982 ◽  
Vol 10 (1) ◽  
pp. 32-33 ◽  
Author(s):  
M. H. JOHNSON ◽  
R. W. H. WALKER ◽  
G. KEIR ◽  
E. J. THOMPSON
Keyword(s):  
New Method ◽  
Polyacrylamide Gels

Preparation of Organometallic and Silyl Complexes of Palladium(II) via [PdCl(SnCl3)L2], where L = PPh3, P(p-Tol)3

2004 ◽  
Vol 69 (7) ◽  
pp. 1464-1471 ◽  
Author(s):  
Małgorzata Noskowska ◽  
Wojciech Duczmal
Keyword(s):  
Reaction Mechanism ◽  
Good Yield ◽  
New Method ◽  
Organohalogen Compound ◽  
Silyl Complexes

A new method for preparing organometallic and monosilyl complexes of Pd(II), based on the interaction between [PdCl(SnCl3)L2] (L = PPh3 or P(p-Tol)3) and appropriate organohalogen compound or chlorosilane has been developed. The products without tin have been obtained with good yield (85-97%). The reaction mechanism has been proposed on the basis of kinetic and chemometric investigations of the reaction between [PdCl(SnCl3)(PPh3)2] and Me2SiCl2.

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