A fourth allele in the plasma esterase-1 (Es-1) system of the domestic fowl

Plasma samples of fowl were analysed by horizontal polyacrylamide gel electrophoresis (pH 9.0). Evidence was presented for the subdivision of an earlier reported esterase-1 allele (Es-1A) into two alleles designated Es-1A1 and Es-1A2. Family data were consistent with the hypothesis that the Es-1 phenotypes were controlled by four codominant, autosomal alleles Es-1C, Es-1A1, Es-1A2 and Es-1B). The White Leghorn samples showed high frequency of Es-1A1 (about 0.7) and also had considerable frequency of Es-1A2 (0.2) and of Es-1B (0.1). The three meat-type breeds studied (White Plymouth Rock, Rhode Island Red and New Hampshire) showed a very high frequency of Es-1B (0.8-1.0).     

Genetic characterization of esterase 28 (ES-28) of the house mouse

The genetics of esterase-28, the major esterase of cauda epididymidis of the house mouse, has been studied after separation by polyacrylamide gel electrophoresis and isoelectric focusing. Four phenotypes are distinguished. Segregation ofEs-28 in two backcross series indicated linkage to Es-1, Es-9, and Es-22. The Es-28 locus was placed into esterase cluster 1 on chromosome 8.This work was supported by the Deutsche Forschungsgemeinschaft.This is communication No. 69 of a research program devoted to the cellular distribution, genetics, and regulation of nonspecific esterases.     

Enzyme polymorphism in the European starling: Study on plasma esterases

In plasma of the European starling (Sturnus v. vulgaris L.), genetic variation was detected by polyacrylamide gel electrophoresis. Two esterase zones, of which one is polymorphic, had already been described earlier. In addition to these two zones (Es-1 and Es-3), we describe a third (Es-2) that also showed variations. The distribution of the phenotypes observed in the Es-2 zone closely fits the expected distribution according to the law of random mating so that the polymorphism of Es-2 is hereditary. This Es-2 locus is postulated to be a codominant system, controlled by two alleles.     

A new variant of the Es-4 locus in the rat

A new variant of kidney esterase in the DK/Nac rat strain is reported. The new esterase was tentatively named ES-4C determined by a third allele of the Es-4 locus of Linkage Group V (LGV). Strain distribution was surveyed using 17 inbred strains, but no strain except for the DK/Nac strain possessed the ES-4C type. Although we surveyed outbred stocks (Jcl: Wistar and Jcl: SD) we could not find rats carrying the ES-4C type. Genetic analysis of the ES-4C type was carried out using mating experiments between DK/Nac and BUF/Nac (ES-4B). The results indicated that the new variant was controlled by the Es-4 locus and it was named the Es-4c allele.     

A new esterase locus, Es-19, in the rat]

A new esterase polymorphism was identified in epididymal homogenates from inbred rat strains by polyacrylamide gel electrophoresis. The inbred rat strains showed either fast (A) or slow (B) bands. Strain distributions of the phenotypes differed from those of other esterase loci. Genetic analyses revealed that the polymorphism is controlled by codominant alleles (Es-19a and Es-19b) and is not linked to linkage groups, I, II, IV, V, VI, XIII of the rat.     

Serum esterase genetics: Identification and hormone induction of the Es-1b esterase in inbred rats

A previously unrecognized esterase from the sera of the appropriate strains of the rat Rattus norvegicus was revealed by a discontinuous polyacrylamide gel electrophoretic technique. This esterase migrated in the albumin region, whereas a previously known major albumin esterase controlled by the Es-2 locus migrated in the postalbumin region when the method was used. The new albumin esterase component which separated from the Es-2 esterase was identified as the product of the Es-1b gene. The new albumin esterase was not detectable in the sera of sexually mature males of the appropriate genotype, because the activity level of this esterase was influenced by sex hormones, especially androgen.     

家鸡血清酯酶遗传多态现象的发育遗传学初步研究

采用垂直平板聚丙烯酰胺凝胶电泳方法随机抽样了后备,初产和产蛋３个阶段的优黄２０００肉种鸡血清酯酶的遗传多态性,结果表明：血清酯酶Ｅｓ－１和Ｅｓ－２均存在遗传多态现象,Ｅｓ－１区检出２－３条多态性酶带,Ｅｓ－２区仅有１条带,表型为带的有或无。     

Heterogeneity of Es-1 esterases in the rabbit (Oryctolagus cuniculus)

Analysis of the Es-1 system in the rabbit with polyacrylamide gel electrophoresis (PAGE) revealed a high degree of individual variation. In the liver the number of esterase bands found in the Es-1 region of the gels ranged from 2 to 16. The results indicate that one locus with three alleles is responsible for all of the esterase bands in the Es-1 region. The most plausible explanation for the observed heterogeneity is that each of the alleles codes for a protein (MW 65,000±2000) that is changed by posttranslational modifications, thus giving rise to two to five monomeric enzymes with esterase activity. Polymerization of these monomers then results in 1–11 dimers. Based on similarities with mouse Es-9, chromosomal homology between rabbit Es-1 and mouse Es-9 is proposed.     

New polymorphisms of esterase 7 and esterase 8 in inbred strains of rats: Tissue expression and linkage studies

Two new esterase polymorphisms (Es-7 and Es-8) were identified in the testis homogenate of laboratory rats, Rattus norvegicus, by using discontinuous gradient polyacrylamide gel electrophoresis. Es-7 expressed two phenotypes: ES-7A (fast) and ES-7B (slow). Es-8, which migrated in the cathodal region rather than the ES-7 region, also expressed two phenotypes: ES-8A (fast) and ES-8B (slow). Linkage tests among Es-2, Es-7, and Es-8 were made from backcross progeny of the mating (LEJ/Hkm × T/Hok)F₁ × LEJ/Hkm. One recombinant in 51 progeny tested was observed between Es-2 and Es-7; however, recombination between Es-2 and Es-8 was not observed in the same progeny. In addition, we show that the esterase polymorphisms of Es-5 in liver homogenate and Es-3 in small intestine homogenate are identical.     

Biochemistry and genetics of esterase-20 (ES-20), a second trimeric carboxylesterase of the house mouse (Mus musculus). I. Purification and characterization of ES-20C1 from male kidney

ES-20 was isolated from male mouse kidney and purified 350-fold by ion-exchange chromatography, isoelectric focusing, and gel filtration. The resultant product was apparently homogeneous by the criteria of polyacrylamide gel electrophoresis and immunodiffusion and represented a major fraction of male mouse kidney esterase. Sodium dodecyl sulfate gel electrophoresis revealed the presence of a single subunit band, molecular weight 59,500; the molecular weight of the native protein was found to be 179,000. Titration of the active site yielded an equivalent weight of about 175,000. The enzyme was further characterized by its kinetic parameters for the hydrolysis of a series of 4-nitrophenyl esters and was classified as a carboxylesterase (EC 3.1.1.1). ES-20C1 bound to concanavalin A, indicating that it was a high-mannose-type glycoprotein; the role of terminal beta-N-acetylglucosamine residues in the carbohydrate side chains for stabilization of the quaternary structure of the trimer was revealed. Extensive biochemical and immunological similarities to ES-9C supported an earlier suggestion that the Es-9c gene product is a component of the ES-20C1 trimer.     

An apparatus for thin layer vertical polyacrylamide gel electrophoresis with discontinuous buffer and gel system

A vertical polyacrylamide gel slab electrophoresis apparatus with a discontinuous gel and buffer system and a running gel of 1 mm in thickness was devised. Using this apparatus, which employs stacking and sieving effects, sharp bands comparable to those of disc electrophoresis were obtained.Furthermore, a new application using detection with ultraviolet method was introduced for isozyme study.     

Esterase. XXI. Es-9, a possibly new polymorphic esterase in Mus musculus genetically linked to Es-2

A so far undescribed gene controlling zone III esterases has been detected by means of disc gel electrophoresis of kidney homogenates from the two inbred mice strains NMRI and SK/Cam. The gene is tentatively designated Es-9, and the two codominant alleles are designated Es-9a and Es-9b. Es-9 esterases are present in many tissues, but, unlike the other zone III esterase (controlled by Es-5), are not found in the serum. Close linkage with the Es-2 gene leads us to map the Es-9 gene on chromosome 8.     

Genetic control of the substrate specificity of sheep plasma arylesterase]

By means of starch gel electrophoresis blood plasma esterases in sheep of different breeds were studied. It is shown that the esterase pattern is a poly-enzyme system consisting of at least three enzymes: arylesterase, carboxylesterase and choline esterase. Postnatal changes of esterase pattern in sheep blood plasma were also studied. Polymorphism on substrate specificities is described, which is expressed in the fact that different arylesterase variants have different affinity to alpha- and beta-isomers of carbone ethers of naphtol. The breeding test suggests that two allelic autosomal genes, reffered to as Es-1a and Es-1b, control the substrate specificity of arylesterase in sheep. The data are discussed in connection with Es-1a and Es-1b gene expression in heterozygous sheep, with the effect of (mosaic) dominance.     

Biochemical Identification of the Two Races of Radopholus similis by Polyacrylamide Gel Electrophoresis

Analysis of proteins of the banana and citrus race of Radopholus similis was carried out by several different types of polyacrylamide gel electrophoresis. These included standard slab gel, SDS slab gel, gradient slab gel, and two-ditnensional slab gel electrophoresis. A major band difference was detected between the two races by slab gel electrophoresis. However, several other poorly resolved but consistent hands of high molecular weight proteins near the gel origin also were considered as diagnostic. Resolution of protein bands was greatly improved by SDS and gradient slab gel electrophoresis, but no differences could be detected among the proteins resolved between the two rares with these techniques. Two-dimensional gels revealed a large number of proteins, but background staining obscured them hindering interpretation. When nematode races were reared on three different host plants, no differences in protein patterns were detected between them, indicating host preferences does not play a role in determining the types proteins occurring in these nematodes.     

Esterase-1: developmental expression in the mouse and distribution of related proteins in other species.

Esterase 1 (Es-1) is a sexually dimorphic 65-kDa glycoprotein present in plasma and other murine tissues able to hydrolyze a variety of esters including fatty acid esters of estradiol. Like most other carboxylesterases, its function is unknown. To gain insight into the function of Es-1 and by analogy other carboxylesterases, we have examined the developmental regulation of Es-1 in the mouse and have looked for the presence of related proteins in the plasma of other species. Northern blot analysis of total RNA from the livers of mice of various ages using a 32P-labeled 470-bp Es-1 cDNA probe showed clear postpartum induction with no detectable Es-1 mRNA in fetal liver. Similarly, immunoblotting after sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an affinity-purified rabbit antibody to Es-1 showed no cross-reacting proteins in the plasma until after birth. Northern blot analysis of total RNA from a variety of adult mouse tissues showed the presence of substantial levels of Es-1 mRNA only in liver with lower levels in kidney, testes, and ovaries. Liver mRNA and plasma protein levels rose in parallel attaining full adult levels between 15 and 20 days of age. When plasma proteins were electrophoresed on 7% polyacrylamide gels under nondenaturing conditions, the antibody to Es-1 recognized a low mobility protein in mouse, rat, human, baboon, guinea pig, bovine, horse, and canine but not in chicken plasma. Consistent with the immunoblotting results, the Es-1 cDNA probe hybridized to restriction fragments from human, monkey, rat, and rabbit as well as mouse genomic DNA but not from chicken DNA indicating conservation of the esterase (or esterase-like) gene in mammalian species. The low mobility antigens in mouse and human plasma appeared also to cross-react with antibodies to human thyroglobulin, although antibodies to human thyroglobulin did not appear to recognize Es-1 under these conditions.     

Esterase-13 of the rat (Rattus norvegicus) and its homology with esterase-3 of the house mouse (Mus musculus)

A new allele of esterase-13 was detected in various laboratory inbred strains of Rattus norvegicus and designated Es-13c. The activity of ES-13 towards a range of chromogenic substrates, inhibitor profile, isoelectric points and retardation coefficients on polyacrylamide gel electrophoresis were determined. The organ specific expression of ES-13 alleles was investigated and it was shown that kidney homogenates contained a factor which modified the liver enzyme banding pattern in vitro. The features of ES-13 from the rat indicated homology between this esterase and ES-3 from the house mouse, Mus musculus domesticus.     

Studies on heterogeneity of the lysine-rich histone from ox pancreas

1. Histone H1 from ox pancrease has been isolated by preparative electrophoresis at pH 2.7 in polyacrylamide slab gel, using the fraction F1 of Oliver et al. (1972, Biochem. J., 129, 349--353) as starting material. 2. The isolated histone H1 showed higher heterogeneity on isoelectric focusing than on polyacrylamide electrophoresis in long gel. The isoelectric points of the main subfractions of histone H1 were at pH 8.0--8.4.     

The genetics of esterase 12 and esterase 13 polymorphisms in the Norway rat

2 new esterase polymorphisms (Es-12 and Es-13) were discovered in haemolysates of wild rats (Rattus norvegicus) by gel electrophoresis. Both loci are probably monomeric. Linkage analysis indicates that neither locus is associated with the esterase cluster in linkage group V.     

Plasma esterase polymorphism in the Bank vole, Clethrionomys glareolus, in Britain

Three hundred and eighty-three Clethrionomys glareolus from 20 localities in England, Wales and Scotland were typed for plasma esterase and a genetic polymorphism was discovered. The esterase was named Es-1. Breeding tests suggested that three alleles were segregating: Es-1^o when homozygous results in complete absence of enzyme activity. The active alleles Es-1^f and Es-1^s code for enzyme variants which migrate more rapidly and less rapidly, respectively, under starch gel electrophoresis. Of these active alleles, Es-1^f is morc common in the north of Britain and Es-1⁸ in the south. A 23-month field study on two areas at Wicken Fen, Cambridgeshire, suggested that animals possessing Es-1^s survived less well at high population densities, perhaps through their being more likely to emigrate.     

Studies on the Molecular Weights of Esterase and Peroxidase Isozymes of Maize

The molecular weights of esterase and peroxidase isozymes of maize seedlings were directly determined by improved polyacrylamide gradient gel electrophoresis. The different isozyme bands developed in polyacrylamide slab gel electrophoresis (uniform gel) were identified in polyacrylamide gradient gel electrophoresis by means of isozyme variants. The molecular weights of esterase isozymes E1, E2, E3F, E3S, a, b, c, named according to isozyme patterns in uniform gel, are <20000, 35200, 33000, 38500, 29900, 28500, 34000 doltons respectively. The molecular weights of peroxidase isozymes PX4F and PX4S are 131000 and 149000 doltons respectively. According to the band location in uniform gel and in gradient gel, some biochemical properties of the isozyme bands and relationships between the isozyme bands were analyzed. The possible errors in the determination of smaller molecular weight isozymes are discussed.