Discontinuous native protein gel electrophoresis

At any stage of growth of a wild-type bakers' yeast, some 20% of the catalatic activity of crude extracts is not precipitable by means of antibody prepared against the typical catalase (catalase T), whose purification and properties have been previously described. Some of this catalatic activity is due to the presence of an atypical catalase (catalase A), a heme protein, with a molecular weight estimated as 170 000 – 190 000, considerably lower than that of the usual catalases (225 000 – 250 000). Preparations of catalase A were found to be homogeneous in the analytical ultracentrifuge and in polyacrylamide gel electrophoresis. Its subunit molecular weight, determined from its iron content, was 46 500, virtually the same as that of the major band obtained in gel electrophoresis in the presence of sodium dodecyl sulfate, suggesting that the native protein is tetrameric. Its specific activity is in the range of those reported for other typical catalases.

Download Full-text

The covalent nature of the human antithrombin III–thrombin bond

Biochemical Journal ◽

10.1042/bj1890481 ◽

1980 ◽

Vol 189 (3) ◽

pp. 481-489 ◽

Cited By ~ 55

Author(s):

M O Longas ◽

T H Finlay

Keyword(s):

Gel Electrophoresis ◽

Antithrombin Iii ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Native Protein ◽

Disulphide Bridge ◽

Terminal Amino Acid ◽

Carboxypeptidase B ◽

Terminal Amino ◽

Covalent Nature

1. Cleavage of the human antithrombin III–thrombin complex with [14C]methoxyamine hydrochloride results in inactive thrombin and 14C-labelled antithrombin III. 2. Discontinuous polyacrylamide-gel electrophoresis of the reduced dissociation fragments of the complex in the presence of sodium dodecyl sulphate reveals two antithrombin III bands that do not resolve during electrophoresis without reduction. The heavy band has the electrophoretic mobility of the native protein. The light band has an apparent mol.wt. that is approx. 4000 less than the molecular weight of native antithrombin III. 3. Treatment of the cleavage products of the complex with carboxypeptidase B yields 1 mumol of arginine, a new C-terminal amino acid, per mumol of thrombin dissociated. The results indicate that during formation of the antithrombin III–thrombin complex, the inhibitor is cleaved at an arginine–X bond; this arginine residue forms a carboxylic ester with the enzyme, while the excised polypeptide remains bound through a disulphide bridge(s).

Download Full-text

Two-dimensional protein gel electrophoresis: A powerful tool in experimental pharmacology and toxicology

Pharmacological Research ◽

10.1016/1043-6618(95)87762-2 ◽

1995 ◽

Vol 31 ◽

pp. 376

Author(s):

L. Aicher ◽

M. Varela ◽

D. Wahl ◽

N.L. Anderson ◽

J.P. Hofmann ◽

...

Keyword(s):

Gel Electrophoresis ◽

Two Dimensional ◽

Protein Gel ◽

Experimental Pharmacology ◽

Pharmacology And Toxicology

Download Full-text

Cold Acclimation and Alterations in Dehydrin-like and Bark Storage Proteins in the Leaves of Sibling Deciduous and Evergreen Peach

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.121.5.915 ◽

1996 ◽

Vol 121 (5) ◽

pp. 915-919 ◽

Cited By ~ 27

Author(s):

Rajeev Arora ◽

Michael Wisniewski ◽

Lisa J. Rowland

Keyword(s):

Cold Acclimation ◽

Gel Electrophoresis ◽

Prunus Persica ◽

Storage Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Subzero Temperature ◽

Protein Gel ◽

Bark Storage Proteins ◽

High Level

Seasonal changes in cold tolerance and proteins were studied in the leaves of sibling deciduous and evergreen peach [Prunus persica (L.) Batsch]. Freezing tolerance [defined as the subzero temperature at which 50% injury occurred (LT50)] was assessed using electrolyte leakage. Proteins were separated by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. Electroblots were probed with anti-dehydrin and anti-19-kD peach bark storage protein (BSP) antibodies. Leaf LT50 decreased successively from -5.8 °C on 18 Aug. to -10.3 °C in the evergreen genotype and from -7.0 °C to -15.0 °C in the deciduous genotype by 14 Oct. Protein profiles and immunoblots indicated the accumulation of a 60- and 30-kD protein during cold acclimation in the leaves of deciduous trees; however, levels of these proteins did not change significantly in the evergreen trees. Immunoblots indicate that the 60-kD protein is a dehydrin-like protein. Gel-electrophoresis and immunoblots also indicated that the 19-kD BSP progressively disappeared from summer through fall in leaves of deciduous peach but accumulated to a high level in bark tissues. A similar inverse relationship was not evident in evergreen peach.

Download Full-text

Effect of Dietary Sorghum Supplemented With Inulin on Ectoparsitic Infection and Protein Gel Electrophoresis of Nile Tilapia (Oreochromis niloticus) Fingerlings

Suez Canal Veterinary Medicine Journal. SCVMJ ◽

10.21608/scvmj.2018.60764 ◽

2018 ◽

Vol 23 (1) ◽

pp. 67-83

Author(s):

smail Essa ◽

Mahi G. ◽

A. Yones ◽

Samar M.

Keyword(s):

Oreochromis Niloticus ◽

Gel Electrophoresis ◽

Nile Tilapia ◽

Protein Gel ◽

Nile Tilapia Oreochromis Niloticus

Download Full-text

A rapid antibody-based test for Sec-2, a marker for the short arm of chromosome 2 of rye (2RS)

Genome ◽

10.1139/g96-125 ◽

1996 ◽

Vol 39 (5) ◽

pp. 1006-1012 ◽

Cited By ~ 5

Author(s):

J. H. Skerritt ◽

R. B. Gupta ◽

J. L. Andrews ◽

F. L. Stoddard ◽

N. K. Howes

Keyword(s):

Gel Electrophoresis ◽

Seed Proteins ◽

Antibody Test ◽

Wheat Chromosome ◽

Chromosome 2 ◽

Wheat Seed ◽

Protein Gel ◽

Seed Population ◽

Selection Of ◽

Rapid Antibody

A simple monoclonal antibody-based screening test has been developed for the presence of translocations of the short arm of chromosome 2 of rye (2RS) with wheat chromosome 2B. 2RS encodes a set of about three polypeptides known as Mr 75 000 gamma-secalins. Use of the antibody test for these secalins enabled screening of several hundred seeds per day. The antibody could readily distinguish 2BL–2RS translocations and 2R substitutions from 1BL–1RS translocations or nontranslocation wheats. Use of the antibody in analysis of segregating progeny for Sec-2 in several wheat backgrounds was successful. Results with a selection of the seed population were checked using protein gel electrophoresis, with 100% correct confirmation. Key words : rye, wheat, seed proteins, translocation, diagnostic test.

Download Full-text