scholarly journals ENZYMES IN HOG KIDNEY HYDROLYZING AMINO ACID NAPHTHYLAMIDES

1966 ◽  
Vol 14 (4) ◽  
pp. 314-325 ◽  
Author(s):  
T. VANHA-PERTTULA ◽  
V. K. HOPSU ◽  
G. G. GLENNER
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Gel Electrophoresis ◽  
Metal Ion ◽  
Total Activity ◽  
Starch Gel Electrophoresis ◽  
Enzyme Preparations ◽  
Starch Gel ◽  
Hydrolysis Of ◽  
Hog Kidney

Hydrolysis of β-naphthylamides of a number of amino acids and dipeptides and of a number of di- and tripeptides by hog kidney homogenate and by fractions obtained by various fractionation procedures has been studied. The substrates were found to be split by a soluble, apparently sulfhydryl-dependent enzyme, and by a particle-bound, metal-activated enzyme. The former constituted only a small part of the total activity. The latter was subfractionated by starch gel electrophoresis into two fractions with identical characteristics. The soluble and particle-bound enzymes differed also in their substrate specificity. The latter enzyme was solubilized, partially purified, characterized by some modifier compounds and compared with enzyme preparations obtained by various fractionation procedures presented by other investigators. Thus enzyme showed ion-determined substrate specificity, i.e., hydrolysis of some of the amino acid naphthylamides was found to be activated by Co++ while the hydrolysis of others was inhibited by the same metal ion.

scholarly journals Organic pyrophosphates as substrates for human alkaline phosphatases

1967 ◽  
Vol 105 (3) ◽  
pp. 1307-1312 ◽  
Author(s):  
R. Helen Eaton ◽  
D W Moss
Keyword(s):  
Gel Electrophoresis ◽  
Human Liver ◽  
Inorganic Phosphate ◽  
Starch Gel Electrophoresis ◽  
Alkaline Phosphatases ◽  
Starch Gel ◽  
Pyrophosphatase Activity ◽  
Small Intestinal ◽  
Hydrolysis Of ◽  
Single Enzyme

1. Purified human liver and small-intestinal alkaline orthophosphatases release inorganic phosphate at appreciable rates from a variety of organic pyrophosphate substrates. 2. The pyrophosphatase action is inhibited by Mg2+ ions at concentrations that activate the hydrolysis of orthophosphate substrates by these enzymes. 3. The results of mixed-substrate experiments, denaturation studies with heat or urea and starch-gel electrophoresis suggest that both orthophosphatase and pyrophosphatase activities are, in each preparation, properties of a single enzyme. 4. Intestinal phosphatase shows greater pyrophosphatase activity relative to orthophosphatase than the liver enzyme.

Comparison of the rates of proteolysis during ripening of Cheddar cheeses made with calf rennet and swine pepsin as coagulants

1974 ◽  
Vol 41 (2) ◽  
pp. 269-282 ◽  
Author(s):  
Margaret L. Green ◽  
P. M. D. Foster
Keyword(s):  
Gel Electrophoresis ◽  
Gluconic Acid ◽  
Gel Filtration ◽  
Total Activity ◽  
Starch Gel Electrophoresis ◽  
Protein Breakdown ◽  
Rate Of Development ◽  
Slow Decline ◽  
Starch Gel ◽  
Acid Lactone

SummaryThe rates of proteolysis during ripening were followed in cheeses made with either calf rennet or swine pepsin and either starter or δ-gluconic acid lactone (GAL) as a replacement for the starter. A gel-filtration column technique and starch-gel electrophoresis were used for analysis, and bacterial counts were made on all samples. Proteolysis was faster in cheeses made using GAL than in those made using starter and also slightly faster in GAL cheeses made with swine pepsin than in those made with rennet. Further, it was considerably slower in starter-containing cheeses made with swine pepsin than in those made with rennet. It is suggested that these differences were due to the much greater rate of development of acidity in cheeses made with GAL than in those made with starter, which resulted in more of the coagulant being incorporated into the curd in an active state. The rate of proteolysis in starter-containing cheeses appeared to follow a characteristic course, being initially slow, then markedly increasing with a later slow decline. It is suggested that the increase in the rate of proteolysis was due to an increase in the total activity of bacterial proteinases released by lysis of the bacteria. Indications were obtained that the coagulants and bacterial proteinases catalysed broadly similar patterns of protein breakdown in cheese, and that medium-sized peptides (mol. wt 9000–14000) were formed as definite intermediates in the process. The results also showed that rennet and swine pepsin remained active for at least 7 months in GAL cheeses, that rennet contributed significantly to proteolysis in starter-containing cheeses, and that swine pepsin was at least extensively inactivated and possibly completely inactivated during cheese-making with starter.

The Separation of Insect Haemolymph Proteins

1964 ◽  
Vol 96 (1-2) ◽  
pp. 110-110
Author(s):  
B. G. Loughton ◽  
P. Rueffel ◽  
H. Stich ◽  
A. S. West
Keyword(s):  
Amino Acid ◽  
Gel Electrophoresis ◽  
Starch Gel Electrophoresis ◽  
Homologous Proteins ◽  
Agar Gel ◽  
Diffusion Technique ◽  
Starch Gel ◽  
Rapid Characterization ◽  
Gel Diffusion

It has been suggested that information on the phylogenetic relationships of genera and species could be obtained by comparing the amino acid sequence in the homologous proteins of different species. This procedure is extremely difficult and time-consuming.However, a relatively rapid characterization of proteins can be obtained by analysing their mobilities with starch-gel electrophoresis and examination of antigenic diversity by the agar gel diffusion technique of Ouchterlony.

scholarly journals The Amino Acid Composition of Hemoglobin. III. A Qualitative Method for Identifying Abnormalities of the Polypeptide Chains of Hemoglobin

Blood ◽  
1964 ◽  
Vol 24 (6) ◽  
pp. 750-756 ◽  
Author(s):  
AMOZ I. CHERNOFF ◽  
NELSON M. PETTIT
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Gel Electrophoresis ◽  
Qualitative Method ◽  
Polypeptide Chain ◽  
Starch Gel Electrophoresis ◽  
Simple Method ◽  
Starch Gel ◽  
Barbital Buffer ◽  
Polypeptide Chains

Abstract A relatively simple method is described which permits the identification of abnormalities of either polypeptide chain of hemoglobin. The procedure is based on the dissociation of hemoglobin by 6 molar urea and starch-gel electrophoresis in a barbital buffer at pH 8.0.

scholarly journals The Fractionation of ?-Histones From Chicken Erythrocyte Nuclei

1964 ◽  
Vol 17 (4) ◽  
pp. 1001 ◽  
Author(s):  
JT Bellair ◽  
OM Mauritzen
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Gel Electrophoresis ◽  
Starch Gel Electrophoresis ◽  
Chicken Erythrocyte ◽  
Exclusion Chromatography ◽  
Starch Gel

Crude IX-histone, obtained from the original histone complex by precipitation of the f3-and y-histones with ethanol, has been shown by starch-gel electrophoresis to contain 13 components. The fractionation of IX-histone by exclusion chromatography on Sephadex G-200 is reported. While none of these components have been obtained pure in the present study, considerable purification of components 2, 3, 4, 5, 8, 9, 10, and 11 has been achieved, and their amino acid composition leaves no doubt that o::-histones represent a muoh larger family of "lysine-rich" proteins than was hitherto supposed.

The tertiary phase of rennin action on αs- and β-caseins

1970 ◽  
Vol 37 (3) ◽  
pp. 437-444 ◽  
Author(s):  
A. M. El-Negoumy
Keyword(s):  
Amino Acid ◽  
Degradation Product ◽  
Gel Electrophoresis ◽  
Phosphate Buffer ◽  
Starch Gel Electrophoresis ◽  
Terminal Amino Acid ◽  
Starch Gel ◽  
Terminal Amino

SummaryCasein, whole αs-casein and β-casein were incubated for 3 and 14 h with crystalline rennin, at pH 6·60 and 36 °C, both in phosphate buffer and in milk dialysate. Products obtained from both systems, comprising 30–83% calciumsensitive (Cas) components, gave similar patterns on starch gel electrophoresis. Whole casein and whole αs-casein were not so soluble in milk dialysate as in phosphate buffer. No significant differences in composition were observed between the Cas and the calcium-insensitive (Ca1) products from the same source.The αs1-component of the Cas product from rennin-treated whole αs-casein had faster gel mobility in comparison to the αs1-component in the Cas product from untreated whole αs-casein. Also, αs1-casein yielded one faster-moving degradation product, while αs2,3,4 appeared unaltered after 14h. The Cas product of rennintreated β-casein also had faster mobility than untreated β-casein and yielded one faster degradation product and several minor ones of slower mobility. Arginine was the only N-terminal amino acid found in the Cas product of both rennin-treated and untreated αs - and β-caseins. The arginine content increased from 3·48 and 4·98 moles/105g to 5·12 and 6·38 moles/105g in the Cas products from rennin-treated β-and αs-caseins, respectively.

scholarly journals Tissue-characteristic forms of glyceraldehyde 3-phosphate dehydrogenase

1970 ◽  
Vol 119 (1) ◽  
pp. 85-88 ◽  
Author(s):  
N. Ressler ◽  
S. Lee
Keyword(s):  
Gel Electrophoresis ◽  
Chloride Solution ◽  
Sodium Chloride Solution ◽  
Total Activity ◽  
Control Mechanisms ◽  
Starch Gel Electrophoresis ◽  
Saturated Sodium Chloride Solution ◽  
Starch Gel ◽  
Glycolytic Control ◽  
Slow Band

Glyceraldehyde 3-phosphate dehydrogenase exists in two different forms in various human tissue preparations. One of them is exhibited, after starch-gel electrophoresis, by a rapidly migrating or `fast' band and the other by a `slow' band. The proportion of the total activity in each of the two forms is characteristic of the type of tissue. A particulate fraction, obtained after centrifugation of homogenates, inhibits the enzyme activity and tends to convert the slow band into a fast one. The conversion is reversible. The fast band can also be converted into the slow one by addition of NAD+ or ADP, or by dialysis against saturated sodium chloride solution. Conversions occur with the purified enzyme as well as with crude homogenates. The relevance of these findings to previous investigations and to glycolytic control mechanisms are discussed.

scholarly journals Investigation into the distribution and properties of histones and other basic proteins in bacteria

1966 ◽  
Vol 101 (3) ◽  
pp. 665-673 ◽  
Author(s):  
JL Leaver ◽  
HJ Cruft
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Gel Electrophoresis ◽  
Basic Protein ◽  
Starch Gel Electrophoresis ◽  
Basic Proteins ◽  
E Coli ◽  
Extraction And Purification ◽  
Starch Gel ◽  
Micro Organisms

1. Methods have been developed for the extraction and purification of bacterial basic proteins. These proteins were initially examined by a micro-method of starch-gel electrophoresis and were then characterized more fully. 2. Although it was found that most of the DNA in both Bacillus megaterium and Escherichia coli must be free from combination with basic protein, there was some evidence that a small portion might be in the form of a typical nucleohistone complex. 3. The ribosomes of both B. megaterium and E. coli were shown to contain approximately 2% of basic protein. On the basis of ultraviolet-absorption curves, partial amino acid analyses and their behaviour on electrophoresis in polyacrylamide gels, it was concluded that some of these ribosomal basic proteins may be extremely similar to typical histones. 4. These results are discussed in relation to those of other authors, and the possible functions of basic proteins present in micro-organisms are considered with reference to those that have already been proposed for the histones of higher organisms.

A QUANTITATIVE METHOD FOR THE DETERMINATION OF CHOLINESTERASE ACTIVITY AFTER STARCH-GEL ELECTROPHORESIS

1966 ◽  
Vol 44 (6) ◽  
pp. 953-955 ◽  
Author(s):  
H. S. Funnell ◽  
W. T. Oliver
Keyword(s):  
Gel Electrophoresis ◽  
Quantitative Method ◽  
Cholinesterase Activity ◽  
Competitive Inhibition ◽  
Starch Gel Electrophoresis ◽  
Final Solution ◽  
Natural Activity ◽  
Starch Gel ◽  
Hydrolysis Of

The method described in this paper is based upon hydrolysis of the starch gel by sodium hydroxide, which releases the dye. The dye is then taken up in an immiscible solvent, and the color determined spectrophotometrically. The method gave good reproducibility in studies both of natural activity and competitive inhibition without requiring any changes in the usual methods. The amount of dye in the final solution was related to the activity towards the dye-producing substrate of the enzyme present in the gel section.

scholarly journals MOUSE ERYTHROCYTE ESTERASES AND AN INTERACTION WITH HEPARIN AS SHOWN BY STARCH GEL ELECTROPHORESIS

1969 ◽  
Vol 17 (2) ◽  
pp. 95-101 ◽  
Author(s):  
McCORMICK TEMPLETON
Keyword(s):  
Young Adult ◽  
Enzymatic Hydrolysis ◽  
Adult Male ◽  
Gel Electrophoresis ◽  
Starch Gel Electrophoresis ◽  
Acetate Esters ◽  
Mouse Erythrocyte ◽  
Starch Gel ◽  
Hydrolysis Of ◽  
Naphthyl Acetate

Esterases from the blood of young adult male C-57 brown mice were separated by starch gel electrophoresis. Enzymatic hydrolysis of the substrates α-naphthyl acetate, naphthyl-AS acetate, α-naphthyl butyrate and acetylthiocholine was studied. Three different esterases were found in erythrocytes. One band hydrolyzed a-naphthyl butyrate but was only slightly active with acetate esters. The fastest migrating bands hydrolyzed acetate faster than butyrate esters, but would hydrolyze both substrates. A third esterase represented in the zymogram by a pair of bands (called "acetate" bands) would hydrolyze only acetate esters. When heparin, or plasma containing heparin, was inserted in a gel so as to overlap an erythrocyte sample, only the acetate bands were altered. When hepanin was inserted cathodal to an erythrocyte sample, the acetate bands are selectively affected by the heparin in such a way as to cause the acetate band to migrate more rapidly toward the anode; other esterase bands were not similarly affected. Acetate bands were not inhibited by physostygmine (1 mM) but their activities diminished with increased concentrations of buffer.

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