Kidney esterases of Mus musculus: Further polymorphism of esterase-6, esterase-9, and a new esterase,esterase-20

The comparison of results obtained by different separation and staining techniques permits the definition of esterase-6 in comparison with esterase-9 and a new esterase, esterase-20. Alleles of Es-6 affect the product's ability to aggregate. Esterase-20 may be an aggregated product of Es-9. The close linkage of Es-6 and Es-9 is confirmed. Homology of esterase-6 with esterases from other mammalian species is also suggested.HRN was supported by the Medical Research Council. This is communication No. 32 of a research program devoted to the cellular distribution, genetics, and regulation of nonspecific esterases.     

Biochemical genetics of rat esterases: Polymorphism,tissue expression,and linkage of four loci

Two alleles at each of four esterase loci in Rattus norvegicus are described with regard to tissue expression, electrophoretic characterization, and genetic linkage. A previously described dominant gene for prealbumin serum esterase is demonstrated to exist as two codominant alleles in the genetically determined absence of the characteristic albumin esterase. The allelic composition of 16 inbred strains for four esterase genes is provided, and the heretofore ambiguous nomenclature of rat esterase genetics is standardized. Linkage of Es-1, Es-2, and Es-3 is demonstrated. Es-2 and Es-3 are tightly linked in that no recombination has been observed in 55 offspring. The same offspring demonstrated 9% recombination between Es-1 and the other two loci.This work was supported by a grant from the Brown-Hazen Fund of Research Corporation.     

Esterase-29 (ES-29): Biochemical characterization and control by two independent gene loci of a testosterone-dependent mouse serum esterase

Biochemistry and genetics of a testosterone-dependent murine serum esterase designated esterase-29 (ES-29) are described. The enzyme was identified after disc electrophoresis and subsequent staining for esterase using -naphthyl acetate as the substrate. It was inhibited by bis-p-nitrophenyl phosphate and was resistant top-chlorophenylsulphonate and hence was classified as carboxylesterase EC 3.1.1.1. The molecular mass was estimated to be about 130 kDa. It was shown that ES-29 is under the control of two independent genes. The first, termed Es-29, is suggested to be a structural locus, linked to the cluster-2 esterase loci on chromosome 8. Three alleles atEs-29, Es-29 ^a, Es-29^b, andEs-29 ^care distinguished, which determine absence (SEG/1), strong activity (BALB/cJ), and low activity (MOLH/Fre), respectively. The second locus, termedMse-1 (serum esterase modifying factor), was found to be closely linked toPre-2 on chromosome 12 and is suggested to be a modifying or regulatory gene. Two alleles were distinguished,Mse-1 ^a(BALB/cJ) andMse-1 ^m(MOL3/JA, CasBgr), which determine whether ES-29 appears as a single band or a double band, respectively.Mse-1 ^mis dominant toMse-1 ^a.This work was supported by the Deutsche Forschungsgemeinschaft. This is communication No. 70 of a research program devoted to the cellular distribution, genetics, and regulation of nonspecific esterases.     

Esterase-17 (ES-17): Characterization and genetic location on chromosome 9 of a bis-p-nitrophenyl phosphate-resistant esterase of the house mouse (Mus musculus)

Genetic variation of a new codominantly inherited esterase, designated ES-17, has been discovered in the house mouse using isoelectric focusing in polyacrylamide gels. The ES-17 A phenotype (three bands; isoelectric points, betweenpH 5.55 andpH 5.90) was found in C57 BL/10Sn. LP/J possessed the Es-17B phenotype (three bands; isoelectric points,pH 5.05–5.55). ES-17 was present in all tissues examined, except for hemolysate and serum, and was most clearly expressed in the small intestine. Because of its reaction toward various substrates and inhibitors, ES-17 has tentatively been classified as acetyl esterase (EC 3.1.1.6). ES-17 was shown to be controlled by the structural locusEs-17, located on chromosome 9. From test-cross data, a gene order ofEs-17-8.7±2.5 map units-Mpi-1-10.2±2.7 map units-Mod-1 was established. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 46). This is communication No. 35 of a research program devoted to the cellular distribution and genetics of nonspecific esterases.     

Es-6, a further polymorphic esterase in the rat

A further polymorphic rat esterase with broad tissue expression and restricted substrate specificity is described and tentatively called Es-6. Inbred rat strains have either fixed allele Es-6^F or fixed allele Es-6^S. Es-6 is not linked to the established esterase cluster consisting of the eight esterase loci Es-1, Es-2, Es-3M, Es-4M, Es-4W, Es-5 (=Es-3W), Es-7, and Es-8 in LG V of the rat or to RT1, Gc, c, a, and h. Esterases with apparently identical biochemical and genetical characteristics are Es-17 of the mouse and Es-A4 of humans.Supported by the Deutsche Forschungsgemeinschaft (Be 352/13 and Gu 105).     

Linkage analyses among five esterase loci in the laboratory rat (Rattus norvegicus)

A cluster of esterase loci has been identified on a segment of a rat linkage group V; however, the linear order of all the loci has not been established. We estimated the recombination frequencies of two locus combinations among five esterase loci (Es-1, Es-2, Es-3, Es-4, and Es-Si) and the linear order of the loci by using three sets of backcross matings: (1) (K:W × IS) × IS, (2) (K:W × IS) × IS, and (3) (SHR × W) × W). The linear order was determined to be Es-1-Es-4-Es-2-Es-3-Es-Si, although the order of Es-2 and Es-4 remains tentative. The sexinfluenced esterase (Es-Si) was demonstrated to be distinct from Es-1 and was proposed to be Es-Si locus with two alleles of Es-Si ^a (positive) and Es-Si ^b (null).This work was partly supported by Grants-in-Aid for Scientific Research, No. 339020 (1978), from the Ministry of Education, Science and Culture, Japan.     

Biochemical genetics of Es-14 (formerly Es-Si) and a new esterase variation,Es-15, of the laboratory rat (Rattus norvegicus): Biochemistry,tissue expression,and linkage to Es-1 in linkage group V

The segregation of rat esterases controlled by loci residing in linkage group V (LGV) has been studied in two backcross series, (LEW/Han × BN/Han)F₁ × LEW/Han and (LEW/Han × LE/Han)F₁ × LEW/Han. Es-14 (formerly Es-Si) was shown to be closely linked to Es-1. A new esterase locus, Es-15, was described which codes for a liver isozyme. The distribution pattern of three alleles at the Es-15 locus is presented for 52 independent inbred strains. Close linkage of Es-15 to Es-14 and to Es-1 was established, proposing the following gene order: [Es-2, Es-10]—[ES-1, ES-14, ES-15]. The esterase loci on LGV are thus separated into two gene clusters, cluster 1 and cluster 2. These conclusions are supported by the strain distribution patterns of the two RI strain series, LXB and DXE.Otto von Deimling was supported by the Deutsche Forschungsgemeinschaft (De 315/2-1, communication No. 56).     

Polymorphism of esterase 11 in Mus musculus,a further esterase locus on chromosome 8

A further esterase, esterase 11, which exhibits a polymorphism detectable by electrophoresis, has been observed in the house mouse, Mus musculus. In 15 inbred strains and two outbred strains, the ES-11A phenotype has been found, composed of two bands of enzyme activity of greater anodal electrophoretic mobility than the two bands of the ES-11B phenotype found in one inbred strain, one wild stock, and 101 wild mice. In F₁ hybrids (IS/Cam×C57 BL/Gr), the phenotype shown corresponds to a mixture of the two parental phenotypes. In backcrosses, ES-11 segregates as an autosomal gene, designated Es-11, closely linked to Es-2 and Es-5 on chromosome 8.This work was supported by the Medical Research Council.     

Genetic control of two esterases of rat plasma (Rattus norvegicus)

Three electrophoretic variants of plasma esterase in the albumin zone, presumably carboxylesterase, have been demonstrated in 250 rats representing a laboratory population of Wistar rats. Electrophoretic variants of the enzyme are believed to be controlled by two codominant alleles at the autosomal locus referred to as Es-2. The variant of carboxylesterase represented by a fast-migrating single band on starch gel electrophoresis is determined by the gene named Es-2 ^a, whereas the slow-migrating variant, represented by two bands, is under control of the allelic gene Es-2 ^b. Animals with Es-2 ^a/Es-2 ^b genotype have three bands of carboxylesterase in the albumin zone. Genetically determined polymorphism of plasma esterase, presumably carboxylesterase, in the prealbumin zone was shown in both laboratory and wild populations of rats. Breeding tests suggest that the gene referred to as Es-1 ^a, responsible for the presence of carboxylesterase in the prealbumin zone, is inherited dominantly, whereas animals homozygous for the allele Es-1 ^b locked this esterase fraction.     

Serum esterase genetics in rats: Two new alleles at Es-2, a new esterase regulated by hormonal factors,and linkage of these loci to the Ag-C blood group locus

Two new alleles at the Es-2 locus are described which determine electrophoretic variants of serum esterases of rats. A new esterase protein is described which is detectable in sera of sexually mature females of the appropriate genotype. Evidence is presented for genetic linkage between the Ag-C blood group locus and Es-1, Es-2, and the locus controlling the sex-influenced protein. Since the Ea-1 blood group locus of mice is linked to four esterase loci, it is suggested that Ag-C is the rat homologue of the mouse Ea-1 locus.This work was supported by U.S.P.H.S. Grants AI-09275, CA-15146, and CA-10097.     

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.     

Genetic identification of non-specific esterases of the mouse cauda epididymidis and description of esterase-28, a new carboxylesterase isoenzyme (EC 3.1.1.1)

Twenty-five (25) electrophoretic bands with esterase activity were distinguished in supernatants of cauda epididymidis of DBA/2J mice. Twenty (20) of these were assigned to 10 genetically defined esterases (ES-1, ES-2, ES-3, ES-6, ES-7, ES-11, ES-14, ES-17, ES-19, ES-22) which were already known from investigations of other mouse tissues. Furthermore, ES-10 was identified in cauda supernatants after isoelectric focussing. A hitherto genetically undefined esterase was assigned to locus Es-28 which was expressed solely in the epididymis. Three phenotypes were distinguished: ES-28A was present in the majority of the inbred strains examined. ES-28B was observed in AKR/Han mice and ES-28C was found in SEG/1 mice.     

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.     

Mapping of theEs-4 locus and linear order of esterase genes (linkage group V; LGV) in the rat

There are three different linear orders of esterase loci of linkage group V (LGV) in the rat (Rattus norvegicus). The first is Es-2-Es-3-Es-1, the second Es-3-(Es-2,Es-4)-Es-1, and the third Es-3-Es-2-Es-1-Es-4. We carried out mating experiments to define the order clearly. Linkage analyses of the four esterase loci, Es-1, Es-2, Es-3, and Es-4, were carried out using two inbred strains carrying different alleles at the four loci. Six locus combinations examined in this study were as follows: Es-1-Es-2, Es-1-Es-3, Es-1-Es-4, Es-2-Es-3, Es-2-Es-4, and Es-3-Es-4. The recombination frequencies of each combination were 6.3, 6.3, 6.3, 5.2, 1.8, and 3.4%, respectively. The first recombination between Es-2 and Es-4 was observed. We propose that the esterase loci of LGV be classified into three clusters according to distances between the loci. The linear order of the four loci is shown to be as follows: [Es-3] (cluster II)-3.4 +/- 2.4%-[Es-4-1.8 +/- 1.7%-Es-2] (cluster III)-6.3 +/- 6.1%-[Es-1] (cluster I).     

Biochemistry and genetics of esterase-20 (ES-20), a second trimeric carboxylesterase of the house mouse (mus musculus). II. A unique recombination reveals ES-20 as a hybrid enzyme

A unique recombination is described between (Es-1, Es-6) and (Es-9, Es-22) within gene cluster 1 of the esterase gene region on chromosome 8 of the house mouse. This recombination established the gene order Es-1--Es-6--(Es-9, Es-22)--Got-2. A further 73 recombinations, from a total of 911 backcrosses, had taken place between cluster 1 and cluster 2. A distance between the clusters of 8.01 +/- 0.90% was calculated; the genes within the clusters appeared more tightly linked than previously assumed. ES-20 behaved anomalously following the recombination within cluster 1: homozygous descendants of the recombinant expressed a new form of ES-20. In vitro incubation of purified ES-6A3 and ES-9A generated ES-20A, revealing this esterase to be a hybrid of different cluster 1 gene products, Es-9 and possibly Es-6. This result satisfactorily accounted for the genetic finding, as well as a range of biochemical properties of ES-20.     

Esr,a second locus in the house mouse controlling esterase-5

Electrophoretic variation characterized by the presence (ES-5B⁺) or absence (ES-5B^–) of esterase-5B in the plasma of the house mouse has been observed. It is suggested that the expression of esterase-5B is controlled by an autosomal locus, Esr, linked to Ldr-1 on chromosome 6, in addition to the presumptive structural locus Es-5, which is located on chromosome 8. A gene order of Lyt-3-Esr-Ldr-1 was determined by two crosses.Supported by the Deutsche Forschungsgemeinschaft (SFB 46).This is communication No. 33 of a research program devoted to the investigation of cellular distribution and genetics of nonspecific esterases.     

Esterase-30 (ES-30) of the house mouse: Biochemical characterization and genetics of a new carboxylesterase isozyme linked to cluster-2 loci on chromosome 8

A new carboxylesterase isozyme (EC 3.1.1.1), designated ES-30, is described in mouse liver. Two phenotypes were distinguished, ES-30A, a possible null type, was found in SPE/Pas and in other lines derived fromMus spretus, and ES-30B was found in BALB/cJ and other laboratory inbred strains. ES-30B is characterized by a distinct electrophoretic band when stained using 5-bromoindoxyl acetate as the substrate. After isolation and purification from other esterases by ion-exchange chromatography and molecular sieving, the molecular mass was estimated by two independent methods to be 62 and 64 kDa, respectively. The activity of ES-30B is higher in adult males than in females and can be stimulatedin vivo by testosterone. The distribution of phenotypes on the progeny of a backcross series suggests a separate locus,Es-30, with the allele a for absence andb for presence of the isozyme. LocusEs-30 is shown to be closely linked toEs-2 and toEs-7 of cluster-2 on chromosome 8. The gene orderEs-9—Got-2—(Es-2, Es-7, Es-30) is suggested. This work was supported by the Deutsche Forschungsgemeinschaft. This is communication No. 72 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.     

Genetic variation and biochemical properties of esterase-18 (ES-18) in the laboratory rat (Rattus norvegicus): A new locus of esterase cluster 2 in linkage group V

A new liver-specific rat carboxylesterase isozyme (EC 3.1.1.1) designated esterase-18 (ES-18) is described. Genetic variation of ES-18 was examined in 93 inbred strains and substrains and a structural locusEs-18 was suggested, coding for either the presence (Es-18 ^a) or the absence (Es-18 ^b) of the isozyme. Linkage studies involving two backcross series revealed thatEs-18 resides in cluster 2 of LGV. No recombination betweenEs-18 and other cluster 2 loci was found in 19 lines of two RI strain sets or in the backcross series.R. K. was supported by the Sonderforschungsbereich 146 (Versuchstierforschung). O.D. was supported by the Deutsche Forschungsgemeinschaft (De 315/2). This is communication No. 65 of a research program devoted to the cellular distribution, regulation, and genetics of nonspecific esterases.     

Genetic control of liver esterase forms in chickens

Six regions of esterase activity designated I to VI were resolved from liver extracts of chickens by horizontal starch gel electrophoresis. These esterases were further characterized on the basis of their substrate affinities and differential responses to various inhibitors.
Genetic variation was found in esterases of region VI which appeared to be ali-esterase. Four phenotypes, A, B, AB and O, were observed. These phenotypes were shown to be controlled by one autosomal locus, designated Es-3 , with alleles Es-3 ^A, Es-3 ^B and Es-3 ^O. This locus is not closely linked to the blood group loci A and B , serum alkaline phosphatase ( Ap ), liver acid phosphatase ( Acp-2 ) and serum esterase ( Es-1 ) loci.