scholarly journals Lens neutral endopeptidase occurs in other bovine and human tissues

1987 ◽  
Vol 248 (3) ◽  
pp. 643-648 ◽  
Author(s):  
K Ray ◽  
H Harris
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Structural Difference ◽  
Double Diffusion ◽  
Neutral Endopeptidase ◽  
Relative Activity ◽  
Polyacrylamide Gel ◽  
Human Tissues ◽  
Banding Patterns ◽  
Bovine Lens

Lens neutral endopeptidase (EC 3.4.24.5) was previously thought to be unique to the eye lens. We report here the finding of a neutral endopeptidase, in a variety of bovine and human tissues, which is very similar both biochemically and immunologically to the lens endopeptidase. SDS/polyacrylamide-gel electrophoresis of partially purified enzyme fractions from various bovine tissues shows the characteristic pattern of at least eight bands with Mr values ranging from 24,000 to 32,000 which was described for the bovine-lens neutral endopeptidase. The relative activity of the enzyme varies from tissue to tissue with lung having the highest activity. Partially purified enzyme fractions from these tissues cross-react with antiserum raised in rabbit against bovine lens endopeptidase showing apparent identity when examined side by side in Ouchterlony double-diffusion tests. The human enzyme also cross-reacts with the antiserum but when tested by double-diffusion against the bovine enzyme the precipitin lines show spurring at the joining edges indicating a structural difference between the human and the bovine enzymes. It was also found by Western blot experiments, after denaturing polyacrylamide-gel electrophoresis of the enzyme, that the polypeptide components of the human and bovine enzymes show somewhat different banding patterns.

Electrophoretic evidence supporting self-incompatibility in Lotus corniculatus

1980 ◽  
Vol 58 (6) ◽  
pp. 712-716 ◽  
Author(s):  
Shirley Dobrofsky ◽  
W. F. Grant
Keyword(s):  
Gel Electrophoresis ◽  
Seed Set ◽  
Polyacrylamide Gel Electrophoresis ◽  
Lotus Corniculatus ◽  
Polyacrylamide Gel ◽  
Self Incompatibility ◽  
Banding Patterns ◽  
Protein Subunits ◽  
Self Pollination

Self-incompatibility, a prefertilization event, and self-sterility, a postfertilization event, have both been suggested as causes for differences in seed set between cross- and self-pollinated florets in Lotus corniculatus L. Ovary protein subunits of selfed, crossed, and unpollinated florets of L. corniculatus cv. Mirabel were studied using polyacrylamide gel electrophoresis. Banding patterns differed for all three conditions. Ovary protein differences were found prior to the time fertilization is known to occur, thereby providing evidence that self-incompatibility is at least partially responsible for the reduced seed set after self-pollination.

Isozymes as markers for identification of tissue culture and greenhouse-grown strawberry cultivars

1991 ◽  
Vol 71 (4) ◽  
pp. 1195-1201 ◽  
Author(s):  
N. S. Nehra ◽  
K. K. Kartha ◽  
C. Stushnoff
Keyword(s):  
Tissue Culture ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Meristem Culture ◽  
Banding Patterns ◽  
Extraction Buffer ◽  
Leaf Tissues ◽  
The Stability ◽  
Strawberry Cultivars

Polyacrylamide gel electrophoresis (PAGE) was used for analysis of isozyme banding patterns of leucine aminopeptidase (LAP), phosphoglucomutase (PGM), phosphoglucoisomerase (PGI), esterase (EST) and 6-phosphogluconate dehydrogenase (6-PGD) in strawberry leaves. The extracts prepared from young leaf tissues using polytron homogenization and an extraction buffer containing 15 mg ml−1 dithiothreitol (DTT) and 10% insoluble polyvinylpyrrolidone (PVP-6755) gave best resolution for these enzymes. The influence of plant age and various growing environments on the stability of isozyme phenotypes was examined. The isozyme banding patterns of 6-PGD were found to vary with the change in growing environment as well as age of the plants. EST produced different banding patterns in greenhouse and tissue culture leaves. However, the isozyme phenotypes of LAP, PGM and PGI remained stable under all the conditions tested. Using a combination of these three stable enzymes, it was possible to distinguish eight strawberry cultivars under both tissue culture and greenhouse conditions. Key words: Fragaria × ananassa Duch., meristem culture, polyacrylamide gel electrophoresis

GENETICS OF GLOSSINA MORSITANS MORSITANS (DIPTERA: GLOSSINIDAE): IV. ELECTROPHORETIC BANDING PATTERNS OF OCTANOL DEHYDROGENASE AND ARGININE PHOSPHOKINASE

1979 ◽  
Vol 111 (11) ◽  
pp. 1307-1310 ◽  
Author(s):  
R.H. Gooding ◽  
B.M. Rolseth
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Aldehyde Oxidase ◽  
Glossina Morsitans Morsitans ◽  
Polyacrylamide Gel ◽  
Single Locus ◽  
Single Individual ◽  
Glossina Morsitans ◽  
Banding Patterns ◽  
Octanol Dehydrogenase

AbstractPolyacrylamide gel electrophoresis of the thoraces of adult Glossina morsitans morsitans Westwood revealed five octanol dehydrogenase (ODH, E.C. 1.1.1.73) phenotypes (and a sixth was predicted) in males and females, two arginine phosphokinase (APK, E.C. 2.7.3.3) phenotypes in males and three APK phenotypes in females. Each enzyme was postulated to be under the control of a single locus; Odh with three alleles on an autosome and Apk with two alleles on the X-chromosome. Allele frequencies were in Hardy-Weinberg equilibrium in our colony, and breeding experiments provided direct evidence for single locus control of each enzyme. The polyacrylamide gel electrophoresis technique described permits determination of banding patterns for xanthine oxidase, aldehyde oxidase, ODH, and APK from a single individual.

scholarly journals THE USE OF SEED PROTEIN BANDING PATTERNS FOR IDENTIFICATION OF PASTURE CULTIVARS

1988 ◽  
pp. 87-91
Author(s):  
M.B. Forde ◽  
S.E. Gardiner
Keyword(s):  
Gel Electrophoresis ◽  
Seed Protein ◽  
Storage Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Subterranean Clover ◽  
Seed Storage Proteins ◽  
Polyacrylamide Gel ◽  
Banding Patterns ◽  
Seed Certification ◽  
Plant Variety

Because of the growing number of pasture cultivars used in NZ and the difficulty of reliably separating cultivars of the same species by morphological characters, seed protein banding patterns have become a useful supplementary means of cultivar identification for the purposes of seed certification and plant variety rights applications. Sodium dodecylsufphate polyacrylamide gel electrophoresis of proteins extracted from ground seed samples produces distinctive patterns of bands representing seed storage proteins of different molecular weights. The procedure can be carried out in two days using viable or dead seed, and the results are not affected by site and season of growth. Although individual seeds of outbreeding species such as perennial ryegrass and white clover produce different banding patterns, the combined population representing the cultivar remains constant unless there has been genetic shift during seed multiplication. Speckes for whrch this procedure is being successfully used include the ryegrasses and fescues, browntop, cocksfoot, bromes, red and white clovers, subterranean clover, serradella and lotus. Even cultrvars as closely related as Nui and Ellett ryegrasses and Huia and Pitau white clovers can be separated by careful work. Because of minor technical differences between runs, all cultivars to be compared must be run on the same gel. Keywords: Seed certification, Plant variety rights, sodium dodecylsulphate polyacrylamide gel electrophoresis.

ISOLATION OF HIGHLY PURIFIED SEX HORMONE BINDING GLOBULIN (SHBG): EVIDENCE FOR MICROHETEROGENEITY

1979 ◽  
Vol 90 (4) ◽  
pp. 737-742 ◽  
Author(s):  
W. Mischke ◽  
H. C. Weise ◽  
D. Graesslin ◽  
R. Rusch ◽  
J. Tamm
Keyword(s):  
Human Serum ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Double Diffusion ◽  
Polyacrylamide Gel ◽  
Sex Hormone ◽  
Sex Hormone Binding Globulin ◽  
Hormone Binding ◽  
Gel Isoelectric Focusing

ABSTRACT Highly purified sex hormone binding globulin (SHBG) was isolated in milligram amounts from a human serum fraction (Cohn IV-4). The final preparation was homogeneous by the criteria of polyacrylamide-gel electrophoresis. Immunological evidence for purity could be given by double diffusion according to Ouchterlony. However, following gel isoelectric focusing highly purified SHBG displayed four different bands, as could be demonstrated by staining as well as by a photoscan of the [3H]5α-dihydrotestosterone-SHBG complex. After incubation with neuraminidase the microheterogeneity of SHBG disappeared and the asialo-SHBG showed only one band.

scholarly journals Identification of Peanut Hybrids Using Microsatellite Markers and Horizontal Polyacrylamide Gel Electrophoresis

2008 ◽  
Vol 35 (2) ◽  
pp. 123-129 ◽  
Author(s):  
S. M. Gomez ◽  
N. N. Denwar ◽  
T. Ramasubramanian ◽  
Charles E. Simpson ◽  
G. Burow ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Leaf Tissue ◽  
Polyacrylamide Gel ◽  
Marker Allele ◽  
Breeding Programs ◽  
Banding Patterns ◽  
Use Of Resources ◽  
Effective Use ◽  
Band Separation

Abstract In peanut hybridization, distinguishing inadvertent selfs from the true hybrids may be difficult. In this study, to differentiate between selfs and hybrids, DNA was extracted from leaf tissue of F1 or F2 plants, and SSR markers were amplified and bands separated by a novel submarine horizontal polyacrylamide gel electrophoresis (H-PAGE). By comparing the resulting banding patterns to those of the parents, 70% of the putative hybrids were shown to be true hybrids on the basis of possessing a marker allele from the male parent. The H-PAGE gels gave better band separation and differentiation of selfed progenies than agarose gels, and were compatible with the common horizontal agarose gel units. This method provides a quick assay to distinguish hybrids from inadvertent selfs, and should result in greater efficiency and more effective use of resources in peanut breeding programs.

Proteins of the Kidney Microvillar Membrane; Analysis by Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis and Crossed Immunoelectrophoresis

1983 ◽  
Vol 65 (5) ◽  
pp. 551-559 ◽  
Author(s):  
Michael T. Abbs ◽  
A. John Kenny
Keyword(s):  
Alkaline Phosphatase ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Human Kidney ◽  
Neutral Endopeptidase ◽  
Dipeptidyl Peptidase ◽  
Dipeptidyl Peptidase Iv ◽  
Polyacrylamide Gel

1. A microvillar fraction was prepared from human kidney cortex. This fraction was seven to 10 times enriched in aminopeptidases N and A, γ-glutamyltransferase, dipeptidyl peptidase IV, neutral endopeptidase and alkaline phosphatase. 2. Dipeptidyl peptidase IV activity of human renal microvilli could be inhibited by di-isopropylphosphorofluoridate and neutral endopeptidase activity by phosphoramidon. 3. Nearly all the activity of aminopeptidases A and N could be removed from the membrane by treatment with papain, but only 19% and 33% of dipeptidyl peptidase IV and γ-glutamyltransferase activities were released under the same conditions. Neutral endopeptidase and alkaline phosphatase were not solubilized by papain. Treatment with elastase gave results similar to papain, except that γ-glutamyltransferase was not released. 4. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis under reducing conditions of microvilli revealed 36 polypeptide bands, 12 of which contained carbohydrate. A band of apparent Mr 130 000 was labelled with [3H]di-isopropylphosphorofluoridate and hence identified as dipeptidyl peptidase IV. 5. Antibodies raised to human kidney microvilli produced 11 precipitates with detergent solubilized proteins and six with papain released proteins. Several of the precipitates were identified histochemically. 6. Microvilli prepared from human kidney are very similar to microvilli from pig and rabbit kidney with respect to enzymology, response to papain treatment, sodium dodecyl sulphate-polyacrylamide gel patterns and immunochemistry.

scholarly journals Proteins of the kidney microvillus membrane. Identification of subunits after sodium dodecylsullphate/polyacrylamide-gel electrophoresis

1976 ◽  
Vol 159 (2) ◽  
pp. 395-407 ◽  
Author(s):  
A G Booth ◽  
A J Kenny
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Apparent Molecular Weight ◽  
Neutral Endopeptidase ◽  
Polyacrylamide Gel ◽  
British Library ◽  
Major Band ◽  
Microvillus Membrane

The proteins of microvilli prepared from pig kidney were analysed by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The typical pattern stained for protein revealed five major bands, four of which also stained for carbohydrate, and about 15 minor bands. For descriptive purposes the bands were designated numerically by their apparent molecular weights (X10(-3). Well-characterized proteins were identified with four of the five major bands. Dipeptidyl peptidase IV, a serine proteinase that may be specifically labelled with di-isopropyl [32P]phosphorofluoridate, was assigned to band 130. Aminopeptidase M was assigned to band 160, though when released from the membrane by a proteinase, this protein comprises three polypeptides each of lower apparent molecular weight than the native enzyme. Neutral endopeptidase can be assigned to band 95 and actin to band 42. The fifth major band (180) is an extrinsic glycoprotein that has not been identified with any microvillus enzyme activity. These four proteins contribute 21% of the microvillus-membrane protein. Kidney microvillus actin was characterized by a variety of properties and was similar to muscle actin. A computer analysis of the gel pattern indicates that it comprises 9.0% of the microvillus protein. Myosin is not present in the microvillus, but another protein associated with band 95, with properties that distinguish it from neutral endopeptidase, was tentatively identified as α-actinin. Alkaline phosphatase was identified as a monomeric polypeptide with an apparent molecular weight of 80000; it is a minor protein of the microvillus and is not discernible as a discrete band in the gel pattern. These and other results permit a model of the organization of the microvillus protein to be suggested. The computer program used has been deposited as Supplementary Publication SUP 50070 (12 pages) at the British Library Lending Division, Boston Spa. Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms given in Biochem. J. (1976) 153,5.

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