HETEROGENEITY OF INSULIN: II. CHROMATOGRAPHY OF INSULIN ON CARBOXYMETHYL CELLULOSE IN UREA CONTAINING BUFFERS

1967 ◽  
Vol 45 (2) ◽  
pp. 221-237 ◽  
Author(s):  
W. W. Dillon ◽  
R. G. Romans
Keyword(s):  
Gel Electrophoresis ◽  
Carboxymethyl Cellulose ◽  
Anomalous Behavior ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Separation ◽  
Minor Components ◽  
Starch Gel ◽  
Starch Gels ◽  
Cellulose Column

Chromatography of insulin, on carboxymethyl (CM) cellulose columns in urea-containing buffers, revealed the presence of at least five components, one major and four minor, in commercial insulin. This fractionation is related to other chromatographic fractionations of insulin and the nature of the minor components is discussed.Insulin which had been incubated in HCl was chromatographed. The behavior of the incubated products on the column showed that two or more amide groups were hydrolyzed simultaneously but at different rates. Because of the anomalous behavior of one of the components, it is postulated that a group other than the amides is also transformed during acid incubation. This component is present in commercial insulin samples.The electrophoretic separation of insulin in urea–starch gels was shown to be due to charge differences on the molecules rather than size differences. Correlation of the components separated on the CM-cellulose column with those separated in the urea–starch gel electrophoresis showed that much smaller charge differences on the molecules are distinguished on the CM-cellulose column than in the gel.

Translation of protamine mRNA in a rabbit reticulocyte cell-free system

1977 ◽  
Vol 55 (2) ◽  
pp. 152-158 ◽  
Author(s):  
Lashitew Gedamu ◽  
Gordon H. Dixon
Keyword(s):  
Gel Electrophoresis ◽  
Carboxymethyl Cellulose ◽  
Starch Gel Electrophoresis ◽  
Stages Of Development ◽  
Free System ◽  
Cell Free System ◽  
Polyadenylated Rna ◽  
Starch Gel ◽  
Three Components

Protamine mRNA isolated from the microsomal and postribosomal supernatant fractions of trout testis in poly A(+) (polyadenylated RNA) and poly A(−) (RNA devoid of poly A(+)) forms (Gedamu, L. &Dixon, G.H. (1976)7. Biol. Chem. 251, 1446–1454 and 1455–1463) was translated in the heterologous rabbit reticulocyte cell-free system; the products were shown to be identical in mobility with authentic protamine by polyacrylamide and starch gel electrophoresis. Chromatography, on carboxymethyl cellulose (Whatman CM-52), of the labelled polypeptide products synthesized in this cell-free system in the presence of poly A(+) and poly A(−) mRNA fractions also showed that [14C]arginine was incorporated into all three protamine components resolved in this system, but there was an unequal and variable incorporation of label into the three components with different preparations of mRNA. These results were interpreted as showing that the population of subcomponents of the protamine mRNA coding for the three different protamine polypeptides varied in batches of trout testis at differing stages of development. In addition, the proportion of mRNA components varied between the poly A(+) and poly A(−) editions of the mRNA, and it appeared that the poly A(−) mRNA fraction might represent the product of deadenylation of an earlier population of poly A(+) mRNA.

INHERITED VARIANTS OF α1-ANTITRYPSIN: A NEW ALLELE PiN

1974 ◽  
Vol 16 (2) ◽  
pp. 297-303 ◽  
Author(s):  
Diane Wilson Cox ◽  
Lynne Celhoffer
Keyword(s):  
Gel Electrophoresis ◽  
Null Allele ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Variants ◽  
Similar Region ◽  
New Variant ◽  
Starch Gel ◽  
Immunofixation Electrophoresis ◽  
Starch Gels ◽  
Antigen Antibody

A new inherited variant of α1-antitrypsin (protease inhibitor or Pi) has been found in five individuals of a family of Welsh origin. The new allele is called PiN, as the α1AT product migrates in acid starch gel between the products of the PiM and PiP alleles. The individuals carrying the PiN allele are all of Pi type MN. The new variant has been compared in several electrophoretic systems with other variants migrating in a similar region by acid starch gel electrophoresis (M, P, S, V, W and X). Acid starch gel and crossed antigen-antibody electrophoresis are most suitable for distinguishing the PiN product. By immunofixation electrophoresis, N has a mobility only slightly different from that of M, however the value of this method can be seen for distinguishing other slow variants which cannot be clearly distinguished on acid starch gels. Twenty-three variants of α1AT are now known. Twenty-two of these are electrophoretic variants and one, the null allele (Pi−), produces no α1AT.

scholarly journals ELECTROPHORETIC SEPARATION OF ESTERASES OF PULMONARY ALVEOLAR CELLS

1970 ◽  
Vol 18 (3) ◽  
pp. 167-177 ◽  
Author(s):  
ROBERT SCHIFF ◽  
MARY ANN BRUNSTETTER ◽  
ROBERT L. HUNTER ◽  
CARROLL E. CROSS
Keyword(s):  
Alveolar Macrophages ◽  
Gel Electrophoresis ◽  
Esterase Activity ◽  
Spectrophotometric Analysis ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Separation ◽  
Alveolar Cells ◽  
Additional Band ◽  
Female Mice ◽  
Starch Gel

Starch gel electrophoresis and vertical flat bed electrophoresis in polyacrylamide gels were used to separate butyrylesterases of pulmonary alveolar macrophages (PAMs) from RF/Al(+) and RF/Al(–) mice. PAM esterases from (+) mice showed five bands in starch and five or six in acrylamide whereas four and three or four bands, respectively, were found in extracts from (–) animals. The additional band or bands present only in positive samples corresponded to the Es-2 prealbumin serum esterase found only in the RF/Al (+) animals. This isozyme was more sensitive to diisopropylfluorophosphate than any other PAM esterase. The serum counterpart was sensitive to the same degree. None of the macrophage esterases were inhibited by eserine sulfate. Spectrophotometric analysis of serum esterase activity indicated a statistically significant increase in female mice from both sublines as compared to their respective males, but did not show a difference between the two sublines. The usefulness of the esterase marker in studies of PAM origin and pathologic conditions is discussed.

Detection of multiple forms of proteolytic enzymes by starch gel electrophoresis

1968 ◽  
Vol 46 (10) ◽  
pp. 1317-1320 ◽  
Author(s):  
E. kaminski ◽  
W. Bushuk
Keyword(s):  
Ionic Strength ◽  
Gel Electrophoresis ◽  
Direct Detection ◽  
Proteolytic Enzymes ◽  
Urea Concentration ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Separation ◽  
Multiple Forms ◽  
Starch Gel ◽  
Sensitive Method

A rapid and sensitive method for the direct detection of multiple forms of proteolytic enzymes by starch gel electrophoresis is described. The location of the enzyme components is identified by the degradation of hemoglobin which is included in the starch gel. This method was used to identify the enzyme components of the 10 commercial proteolytic enzymes bromelain, chymotrypsin, ficin, papain, pepsin, pronase B, protease, proteinase, and two preparations of trypsin. The effects of urea concentration and the ionic strength of aluminium lactate buffer were also examined. The best results were obtained with 3 M urea and with an ionic strength of 0.1 for the lactate buffer. It was observed that the number of enzyme components decreased with increasing concentrations of urea or increasing ionic strength of lactate buffer. The number of enzyme components did not always correspond to the number of protein bands. Self-digestion occurred in some of the protein bands in the starch gel after electrophoretic separation of the proteolytic enzymes.

scholarly journals An Improved Chromatographic Separation of Wheat Gluten Proteins

1965 ◽  
Vol 18 (1) ◽  
pp. 193 ◽  
Author(s):  
CW Wrigley
Keyword(s):  
Column Chromatography ◽  
Chromatographic Separation ◽  
Gel Electrophoresis ◽  
Carboxymethyl Cellulose ◽  
Wheat Gluten ◽  
Starch Gel Electrophoresis ◽  
Gluten Proteins ◽  
Electrophoretic Patterns ◽  
Starch Gel

Gel electrophoresis is at present the best procedure for the analytical fractiona-tion of wheat gluten proteins. Procedures more suitable for preparative studies, such as gel ffitration and column chromatography, have provided only partial fractionation on the basis of gel electrophoresis (Graham 1963; Lee et al. 1963; Wright, Brown, and Bell 1964). Gel electrophoretic patterns of fractions resulting from the carboxymethyl cellulose (CMC) fractionation of Simmonds and Winzor (1961) have been published by Graham (1963) and by Lee et al. (1963). These results showed that each fraction was heterogeneous, with considerable overlapping of electrophoretic bands from neighbouring fractions, and that fractions eluted late in the chromatogram were contaminated because of tailing of earlier fractions. The modification of the Simmonds and Winzor procedure described below results in an improved fractionation of gluten proteins as judged by starch-gel electrophoresis.

Studies on the relation between plasma antithrombin and heparin-cofactor

1968 ◽  
Vol 46 (3) ◽  
pp. 347-350 ◽  
Author(s):  
F. C. Monkhouse ◽  
Susan Milojevic
Keyword(s):  
Gel Electrophoresis ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Fold Increase ◽  
Significant Loss ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Patterns ◽  
Starch Gel ◽  
Cofactor Activity ◽  
Cellulose Column

A method for the preparation of purified plasma antithrombin and heparin-cofactor is described. The method involves adsorption by aluminium hydroxide, separation on a DEAE-cellulose column by means of a graded salt concentration, and vertical curtain electrophoresis. A 100-fold increase in the specific activity of antithrombin and a 30-fold increase in the specific activity of heparin-cofactor have been achieved. In spite of the increased purification, no separation of the two activities was achieved. When a highly purified fraction was subjected to starch-gel electrophoresis for 16–18 h and then eluted from the gel, there was significant loss of heparin-cofactor activity but not of antithrombin activity. The electrophoretic patterns of the recovered proteins were not altered.

Proteolysis detection in milk: IV. Starch-gel electrophoresis and formol titration

1975 ◽  
Vol 42 (2) ◽  
pp. 277-283 ◽  
Author(s):  
H. S. Juffs
Keyword(s):  
Gel Electrophoresis ◽  
Bacterial Count ◽  
Raw Milk ◽  
Total Bacterial Count ◽  
Starch Gel Electrophoresis ◽  
Quality Indices ◽  
Bacteriological Quality ◽  
Starch Gel ◽  
Starch Gels

SummaryStarch-gel electrophoresis (SGE) and formol titration methods for detecting proteolysis in cold-stored raw milk have been studied to establish their value as quality indices. When examined by SGE, the first evidence of proteolysis in raw milks stored at 5°C was the formation of para-κ-casein. However, this fraction could not be detected on the starch gels until the total bacterial count (TBC) exceeded 107/ml. The SGE method appeared more reliable than the previously discussed tyrosine value method. Formol titration did not appear to have any application in the screening of cold-stored raw milks with TBC < 107/ml, but would detect some milks of poorer bacteriological quality.

Fractionation of Plasminogen by Starch Gel Electrophoresis

1964 ◽  
Vol 12 (01) ◽  
pp. 126-136 ◽  
Author(s):  
Karl H. Slotta ◽  
J. D Gonzalez
Keyword(s):  
Gel Electrophoresis ◽  
Room Temperature ◽  
Broad Band ◽  
Caproic Acid ◽  
Starch Gel Electrophoresis ◽  
Full Activity ◽  
Veronal Buffer ◽  
Starch Gel ◽  
Alkaline Range

SummaryWhen urea or ε-amino caproic acid were used as solublizing agents for plasminogen in electrophoretic experiments, only one broad band of the proenzyme was obtained on acetate cellulose, in starch block, and in acrylamide gel. In starch gel electrophoresis, however, both forms of plasminogen – the native or euglobulin and Kline’s or Pseudoglobulin plasminogen – separated into six bands. These migrated toward the cathode at room temperature in borate or veronal buffer in the alkaline range and showed full activity in fibrinagar-streptokinase plates.

scholarly journals THE INHERITANCE OF ENZYME VARIANTS FOR TYROSINE AMINOTRANSFERASE, NADP-DEPENDENT MALATE DEHYDROGENASE, NADP-DEPENDENT ISOCITRATE DEHYDROGENASE, AND TETRAZOLIUM OXIDASE IN TETRAHYMENA PYRIFORMIS, SYNGEN 1

Genetics ◽  
1973 ◽  
Vol 74 (4) ◽  
pp. 595-603
Author(s):  
D Borden ◽  
E T Miller ◽  
D L Nanney ◽  
G S Whitt
Keyword(s):  
Detailed Analysis ◽  
Malate Dehydrogenase ◽  
Isocitrate Dehydrogenase ◽  
Gel Electrophoresis ◽  
Tetrahymena Pyriformis ◽  
Tyrosine Aminotransferase ◽  
Breeding Program ◽  
Starch Gel Electrophoresis ◽  
Enzyme Locus ◽  
Starch Gel

ABSTRACT The isozymic patterns of tyrosine aminotransferase, NADP malate dehydrogenase, NADP isocitrate dehydrogenase, and tetrazolium oxidase were examined by starch-gel electrophoresis in Tetrahymena pyriformis, syngen 1. The genetics of the alleles controlling these enzymes was studied through a breeding program. Each enzyme locus was shown to assort vegetatively, as do other loci in this organism. A detailed analysis of the assortment process for the tyrosine aminotransferase locus indicated that the rate of stabilization of heterozygotes into pure types was essentially identical to previously-reported rates for other loci.

Sign in / Sign up

Export Citation Format

Share Document