Isoelectric focusing of red cell phosphoglucomutase (E.C.:2.7.5.1) at the PGM1 locus in a French-Canadian population

Summary Phosphoglucomutase₁ (PGM₁) polymorphism was studied in a French-Canadian population of Québec city, Canada by means of a low voltage (max 500 V) isoelectric focusing (IEF) procedure on vertical polyacrylamide gel slabs. Frequencies of the four common PGM₁ genes estimated from the phenotype distribution in 308 unrelated individuals were PGM ₁ ¹⁺ , 0.61 (±0.02); PGM ₁ ^1- , 0.13 (±0.01); PGM ₁ ¹⁺ , 0.61 (±0.02); PGM ₁ ^1- , 0.18 (±0.02); and PGM ₁ ¹⁺ , 0.61 (±0.02); PGM ₁ ^1- , 0.08 (±0.01). The segregation patterns observed in 154 families, which included 31 different mating types and 353 children, confirmed a Mendelian inheritance of four autosomal genes. The distribution of the PGM₁ phenotypes observed or expected in a Hardy-Weinberg equilibrium was compared with that of other populations. A significant (P<0.001) difference was found between the Québec population and a Black population from Keneba, Gambia, West-Africa.     

Rare phenotypes of the phosphoglucomutase locus 1 detectable by isoelectric focusing on Cellogel

Summary The rare phenotypes PGM₁, determined by alleles PGM ₃ ¹ , PGM ₄ ¹ , PGM ₆ ¹ , and PGM ₇ ¹ were examined by starch gel electrophoresis and cellulose acetate gel isoelectric focusing and were compared with the commonest phenotypes of PGM₁.The frequencies of the rare genes found in the Polish populations were as follows: in Lublin, PGM ₃ ¹ =0.0002, PGM ₄ ¹ =0.0005, PGM ₆ ¹ =0.0010, and PGM ₇ ¹ =0.0005; in Wroclaw, PGM ₃ ¹ =0.0000, PGM ₄ ¹ =0.0005, PGM ₆ ¹ =0.0007, and PGM ₇ ¹ =0.0002.The results suggest that the F and S type variants of the genes PGM ₄ ¹ and PGM ₇ ¹ probably do not occur. It is still possibile that F and S variants exist for the genes PGM ₃ ¹ and PGM ₆ ¹ .     

An improved method of typing hair sheath cells using the PGM3 locus following starch gel electrophoresis

Summary The PGM₃ locus, like the PGM₁ locus, is shown to be easily demonstrated in hair sheath cells using starch gel electrophoresis. The discriminating power of the total system (PGM₁ and PGM₃) on starch gel electrophoresis closely approaches that observed by isoelectric focusing of the PGM₁ locus. Family studies of the PGM₃ locus variants in hair sheath cells confirm that the alleles responsible are inherited in a Mendelian fashion independent of the PGM₁ locus.     

Heterogeneity of phosphoglucomutase

A survey of the isoenzyme patterns of phosphoglucomutase (PGM) as demonstrated by starch gel electrophoresis has been undertaken in different species. Seven mammals, two birds, one reptile, two amphibians, four fish, and four invertebrates were studied, and in some cases several tissues were examined. In all cases the predominant enzyme present was a group of electrophoretically related enzymes which are believed to all represent expression of the PGM₁ allele. In some species, examples of other groups of isoenzymes were encountered, presumably representing other genetic loci, corresponding to PGM₂ and PGM₃. These were always less in amount. The PGM from the chicken was unique in the survey in that its mobility changed with storage. A somewhat similar although not identical change could be produced by addition of PCMB to the partially purified enzyme. Both altered enzymes, i.e., that resulting from storage and that produced by addition of PCMB, were more heat labile.Supported by a grant from the National Foundation for Neuromuscular Diseases, and by the Mental Retardation and Human Development Research Program (1 PO1 HD 03773-01).     

Population studies on human phosphoglucomutase-1 thermostability polymorphism

Summary The electrophoretic and thermostability polymorphisms of the PGM ₁ locus were examined in about 700 Czechoslovakians (Prague) and 3000 Italians. The Italian sample consisted of individuals from Pavia (Northern Italy), Viareggio and Rome (Central Italy) and Naples (Southern Italy). The eight PGM ₁ alleles, PGM ₁ ^1Str , PGM ₁ ^1Sts , PGM ₁ ^1Ftr , PGM ₁ ^1Fts , PGM ₁ ^2Str , PGM ₁ ^2Sts , PGM ₁ ^2Ftr , PGM ₁ ^2Fts , have been considered as combinations of mutations at three different sites, 1/2 S/F and tr/ts, within the PGM ₁ gene and their frequencies discussed in terms of linkage disequilibrium between these sites. All pairwise differences between the samples were significant except for Pavia-Viareggio and Viareggio-Rome. The frequencies of the PGM ₁ ^ts alleles have been found to range from 0.0981 (Prague) to 0.0546 (Naples) and can be ordered according to a North-South cline.This paper is dedicated to Professor Giuseppe Montalenti in occasion of his 80th birthday     

Genetic markers in Malaysians: Variants of soluble and mitochondrial glutamic oxaloacetic transaminase and salivary and pancreatic amylase,phosphoglucomutase III and saliva esterase polymorphisms

Summary Malaysians of Malay, Chinese, and Indian ancestries were electrophoretically phenotyped for Amy₁ and saliva esterase region 1(Set-1) from saliva, Amy₂ from plasma, soluble and mitochondrial GOT and PGM ₃ from leukocyte and placenta. Kadazans and Bajaus, the indigenous people of Sabah, East Malaysia were surveyed for Amy₂. Three types of variants were observed for Amy₁, one type for Amy₂. Only Indians were found to be polymorphic for Amy₁. Two GOT _s 2-1 and three GOT _m 2-1 variants were found among 281 Chinese while three GOT _m 2-1 variants were found among 311 Malays.Malaysian Malays, Chinese, and Indians were found to be polymorphic for Set-1 and PGM ₃. The gene frequencies in Malays are Set-1F=0.601±0.021, Set-1S=0.399±0.021; PGM ₃ ¹ =0.788±0.020, PGM ₃ ² =0.212±0.020; in Chinese Set-1F=0.497±0.028, Set-1S=0.503±0.028; PGM ₃ ¹ =0.745±0.024, PGM ₃ ² =0.255±0.024; in Indians, Set-1F=0.449±0.031, Set-1S=0.551±0.031; PGM ₃ ¹ =0.755±0.029, PGM ₃ ² =0.245±0.029.     

Effects of various metabolites on two phosphoglucomutase allozyme activities from Drosophila melanogaster

The effects of various metabolites on the two most common phosphoglucomutase allozymes (PGM^A and PGM^B) in Drosophila melanogaster have been investigated in vitro. 2,3-Diphosphoglycerate (2,3DPG) inhibited PGM^A and PGM^B to the same degree in the presence of 25 µM glucose-1,6-diphosphate (G1, 6P₂). However a higher concentration of G1,6P₂ partially reversed the inhibition of PGM^A exerted by 2,3DPG, so that in the presence of 150 µM G1,6P₂ the inhibition of PGM^A was half that of PGM^B at pH 6.0. Glycerol-3-phosphate (G3P) had no significant effect at pH 7.4 but exerted an activating effect at pH 6.0 which was more pronounced in the case of PGM^B. ATP, citrate, and fructose-1, 6-diphosphate (F1,6P₂) inhibited both PGM^A and PGM^B. The differences found in vitro between these two allozymes can have a significant impact on in vivo function and, therefore, on the maintenance of PGM polymorphism in experimental populations of D. melanogaster studied in the laboratory.     

Red cell enzymes of the Pongidae

Summary The red cell enzymes acid phosphatase, adenylate kinase, adenosine deaminase and phosphoglucomutase were analyzed by horizontal starch gel electrophoresis in 43 members of the family Pongidae: Pongo pygmaeus (n=10), Gorilla g. gorilla (n=8), Pan troglodytes (n=22) and Pan paniscus (n=3).In all the Pongidae a red cell acid phosphatase zymogram corresponding to the phenotype B in man was found. The adenylate kinase corresponded to the human phenotype AK 1. All the Pongidae showed the same homozygous adenosine deaminase phenotype which was different from the zymograms in man and was designated ADA ape. In all Pongidae the allele PGM ₁ ¹ was present, in addition in Gorilla g. gorilla a second allele was demonstrated, PGM ₁ ^Go . In Pan troglodytes a second allele, PGM ₁ ^Pan was recognized. In Pongo pygmaeus and Gorilla g. gorilla the PGM₂ patterns differed in their migration rates from PGM₂ 1 in man. In one individual of the species Pan troglodytes a PGM₂ zymogram was found resembling the heterozygous phenotype PGM₂ 3–1, PGM ₂ ¹ PGM ₂ ³, (type Palmer) in man. In all the other individuals of the species Pan troglodytes and in those of the species Pan paniscus the PGM₂ zymogram corresponded to the phenotype PGM₂ 1 in man.

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Zusammenfassung Bei 43 Vertretern der Familie Pongidae, Pongo pygmaeus (n=10), Gorilla g. gorilla (n=8), Pan troglodytes (n=22) und Pan paniscus (n=3), wurden die Erythrocytenenzyme saure Phosphatase, Adenylatkinase, Adenosindeaminase und Phosphoglucomutase mit der horizontalen Stärkegelelektrophorese analysiert. Bei allen Pongiden fanden wir eine saure Phosphatase, die dem Phänotyp B des Menschen entsprach, und eine Adenylatkinase, die dem Phänotyp AK 1 des Menschen glich. Alle Pongiden besaßen das gleiche, einem homozygoten Phänotyp entsprechende Adenosindeaminase-Zymogramm, das sich von den Zymogrammen des Menschen unterschied; wir bezeichnen diesen Phänotyp mit ADA ape. Bei allen Pongiden kommt das Allel PGM ₁ ¹ vor, bei Gorilla g. gorilla zusätzlich ein zweites Allel, PGM ₁ ^Go , und bei Pan troglodytes ein zweites Allel, PGM ₁ ^Pan . Die PGM₂-Zymogramme von Pongo pygmaeus und Gorilla g. gorilla unterschieden sich in ihrer elektrophoretischen Wandergeschwindigkeit vom Phänotyp PGM₂ 1 des Menschen. Bei einem Individuum der Species Pan troglodytes fanden wir ein heterozygotes PGM₂-Zymogramm, das an den heterozygoten Phänotyp PGM₂ 3–1, PGM ₂ ¹ PGM ₂ ³ (Typ Palmer) des Menschen erinnerte, bei allen übrigen Individuen der Species Pan troglodytes und bei denen der Species Pan paniscus ein homozygotes PGM₂-Zymogramm, das dem Phänotyp PGM₂ 1 des Menschen entsprach.

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Supported by the Deutsche Forschungsgemeinschaft.     

Results of haptoglobin types investigations in Hungary

Summary This is report about the PGM and PGD isozymes that are present in sperm cells. 4 isozymes that are controlled by the PGM₁ alleles can be detected regularly. Only one isozyme is identified as a product of the PGM ₂ ¹ gene. The isozymes that are controlled by the PGM₃ locus are detected at regular intervals. PGD patterns differ only slightly from those of red cells.

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Zusammenfassung Es wird über die PGM- und PGD-Isoenzyme, die in Spermatozoen vorkommen, berichtet. Als Produkte der PGM₁-Allele lassen sich regelmäßig 4 Isoenzyme erkennen. Nur ein Isoenzym ist erkennbar, welches dem PGM ₂ ¹ -Gen zugeordnet werden kann. Weiterhin lassen sich regelmäßig die Isoenzyme des PGM₃-Locus erkennen. Die PGD-Muster in Spermapherogrammen ähneln denen aus Erythrocytenhämolysaten.

     

Differential function of the phosphoglucomutase isozymes PGM1 and PGM2

Summary A total of 13 metabolites thought to be possibly inhibitory were tested for their influence on PGM isozyme activities, each at several different concentrations. The analysis of statistical significance was based on enzyme activities obtained by densitometric measurements of starch gels. Five of the substances were found to inhibit PGM activity, three of which definitely and a further one probably led to a significantly stronger inhibition of the isozymes of the PGM ₂ locus than of PGM ₁ isozymes. They are (1) fructose-1,6-diphosphate, (2) adenosine triphosphate, (3) citrate, and (4) possibly 2,3-diphosphoglycerate. Thus, PGM ₁ isozymes proved to function better in hard or perhaps marginal metabolic conditions. Related evolutionary aspects are discussed.     

Investigation of PGM13, PGM16, and PGM17 variants by isoelectric focussing. Evidence for new subtypes of the PGM 1 3 and PGM 1 7 alleles

Summary A total of 345 haemolysates previously phenotyped by starch gel electrophoresis and known to contain the products of the PGM ₁ ³ , PGM ₁ ⁶ , and PGM ₁ ⁷ alleles have been analyzed by thin layer polyacrylamide gel isoelectric focussing in the pH range 5–7. Two common subtypes, 3+and 3-, of the PGM ₁ ³ allele have been found in a number of Pacific populations. A single form of the PGM ₁ ⁷ allele was observed in the Western Caroline Islands. In contrast, one of two Indian PGM₁7 variants focussed to a different position when compared with the form found at polymorphic frequency in the Western Caroline Islands. Only one type of the PGM ₁ ⁶ allele was detected during the present investigation.     

Isoelectric focusing of human red cell phosphoglucomutase (PGM1). Phenotype distribution in the population of Tuscany and two hereditary variants

Summary Phosphoglucomutase (PGM₁) phenotypes were determined in a population sample of Tuscany, Italy, by isoelectric focusing. The frequencies observed for the four alleles are: PGM ₁ ¹⁺ =0.6012, PGM ₁ ^1- =0.1059, PGM ₁ ²⁺ =0.2495, PGM ₁ ^2- =0.0434. Two variants were detected and it was possible to study the parentage of both of them. The pedigree of the propositus of the first variant shows that the variant occurs in combination with the common alleles PGM₁ 1+ and PGM₁ 2+ and that it has an autosomal dominant inheritance. The second variant has been shown to be a product of the PGM₂ locus, although its PAGIF pattern is included between 2- and 1+ isoenzymes.     

Substrate affinity in PGM1, PGM2, and PGM3 isozymes

Summary An easy method for routine detection of PGM₁, PGM₂, and PGM₃ isozymes is given. Differences in substrate affinity are discussed. Gene products pgm₁ can be differentiated from gene products pgm₃ by cofactor requirement.     

Polymorphic Analysis of the Human Phosphoglucomutase-3 Gene Based on Mismatched PCR–RFLP Technique

Polymorphic analysis of human phosphoglucomutase-3 (PGM₃) has been carried out from the level of the gene product. Due to a weak zymogram, leading to ambiguity in phenotyping, information on the PGM ₃ locus has rarely been reported. In this study, the missense mutation G1396A, confirmed to underlie common phenotypes of PGM₃, was identified by performing mismatched PCR–RFLP. Population data on the PGM ₃ locus was also obtained for the first time in China. The allele frequency distribution was PGM ₃ *1 = 0.625, PGM ₃ * 2 = 0.375, and no deviation from Hardy–Weinberg equilibrium was observed. The application of the information in both genetics and forensic medicine demonstrated that the polymorphism information content was 0.5163, heterozygosity 0.4872, power of discrimination 0.5986, and probability of paternity exclusion 0.1794. Polymorphic analysis of the locus at the DNA level will also provide significant data for disease susceptibility and linkage analysis.     

Beitrag zur Genetik der Erythrocyten-Phosphoglucomutase

Summary Blood samples from 515 unrelated individuals and from 41 families with 80 children living in Hessen (Germany) were examined for erythrocyte phosphoglucomutase (PGM₁) polymorphism. The gene frequences especially fit well with those obtained in Berlin and Danmark. The family studies verify the supposed way of heredity without any exception.

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Direktor: Prof. Dr. W. Spielmann     

Population genetic studies in three Chinese minorities

Three minority ethnic groups from China (Mongolians, Koreans, Zhuang) were examined with respect to the genetic markers GLO, GPT, ACP, ESD, 6-PGD, PGM₁ subtypes, C3, and TF. Significant variations were noted for the gene frequencies of GLO, GPT, ESD, sub PGM₁ between Zhuang and Mongolians; for GPT, ACP, ESD, sub PGM₁ between Zhuang and Koreans; and for GLO between Mongolians and Koreans.     

Separation of bovine spermatozoa proteins using 2D-PAGE revealed the relationship between tektin-4 expression patterns and spermatozoa motility

Poor semen quality has long been associated with bull infertility. However, the molecular basis in spermatozoa cells underlying the mechanisms of bull infertility remain unknown. The purpose of this study was to determine whether there is any protein in bovine spermatozoa related to semen quality. Semen samples from 18 Brahman bulls, 3 to 10 yrs of age, were assessed for semen quality in terms of spermatozoa motility and spermatozoa morphology. Spermatozoa extracts were separated using 2D-PAGE followed by staining with Coomassie blue. At least one duplicate gel was performed for each sample. Each gel was scanned with an ImageScanner System and analyzed for spots by ImageMaster 2D platinum software. The related protein spot(s) with semen quality was cut from the gel and identified by LC MS/MS. The results showed that at least 600 protein spots were detected in the spermatozoa extracts of the Brahman bulls. Of all these spots, there were 3 of 56 kDa at pI 6.4, 6.6 and 6.8 (Z₁, Z₂ and Z₃, respectively) that clearly showed different expression pattern among 18 Brahman bulls. Of 18 bulls (a) five showed the presence of spot Z₁ and Z₂ (pattern A) (b) one of spot Z₃ (pattern B) (c) five of spot Z₂ and Z₃ (pattern C) (d) one of spot Z₁ (pattern D) and (e) six of spot Z₂ (pattern E). Identification of spot Z₁, Z₂ and Z₃ by LC MS/MS had a similar result as matched to the tektin-4 protein of Bos taurus with a respective score of 171, 557 and 591. The statistical analysis of the 56 kDa protein patterns, tektin-4, indicated a significant effect on spermatozoa motility (P < 0.05) albeit non-significant on spermatozoa morphology. The bulls which showed pattern A had a higher percentage of spermatozoa motility than pattern E (P < 0.05) and not different from pattern C (P > 0.05). The statistical analysis also revealed that the presence of spot Z₁ had an effect on the percentage of spermatozoa motility (P < 0.01), whereas the presence of spot Z₂ and Z₃ had no effect (P > 0.05). The correlation coefficient between the relative protein content of spot Z₁ and the percentage of spermatozoa motility was 0.49. Our study demonstrates that the expression patterns of tektin-4 were a proxy for an effect on spermatozoa motility and consequently bull infertility. It may be that these protein patterns can be used as markers for improving bovine reproduction.     

Characterisation of the isoenzymes of phosphoglucomutase (PGM) determined by the first (PGM1) and second (PGM2) locus observed by isoelectric focusing

Summary The existence of four alleles of phosphoglucomutase (PGM₁) in human red cell lysates has previously been demonstrated by isoelectric focusing (Bark et al., 1976; Kühnl et al., 1977; Sutton and Burgess, 1978). Experiments are now described in which the position of each of the first-locus (PGM₁) and second-locus (PGM₂) isoenzymes is defined, thus extending and confirming the original proposal made by Bark et al.     

Problematischer Mutter-Kind-Ausschluß mit PGM1

Summary An apparently false exclusion of maternity (PGM₁1 versus PGM₁2) indicates the possibility of an additional allele of the PGM₁ locus, which cannot be detected by the usual electrophoretic methods.

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Direktor: Prof. Dr. G. G. Wendt

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Direktor: Prof. Dr. Dr. H. Ritter

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Polymorphism of erythrocyte phosphoglucomutase,adenylate kinase and adenosine deaminase in northern Thailand

Summary Phenotypes of the erythrocyte enzymes phosphoglucomutase (PGM) (n-587), adenylate kinase (AK) (n=695), and adenosine deaminase (ADA) (n=616) were determined by horizontal starch gel electrophoresis in Thai subjects from norther Thailand, mainly from the provinces of Chiang Mai and Lamphun. The following gene frequencies were calculated: PGM ₁ ¹ 0.7385 PGM ₁ ² 0.2487 PGM ₁ ⁶ 0.0102 PGM ₁ ⁷ 0.0026, AK ¹ 0.9950 AK ² 0.0050, ADA ¹ 0.9180 ADA ² 0.0820.The regular, apparently autosomal transmission of the PGM ₁ ⁶ and PGM ₁ ⁷ alleles was demonstrated in 7 families revealing sufficient data.

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Zusammenfassung Die Phänotypen der Erythrocytenenzyme Phosphoglucomutase (PGM) (n=587), Adenylatkinase (AK) (n=695), and Adenosindeaminase (ADA) (n=616) wurden mittles horizontaler Stärkegelelektrophorese bei Thailändern aus Nordthailand, hauptsächlich aus den Provinzen Chiang Mai und Lamphun, bestimmt. Auf Grund der Ergebnisse wurden die in der englischen Zusammenfassung angegebenen Genfrequenzen berechnet. Die regelmäßige, anschinend autosomale Vererbung der Allele PGM ₁ ⁶ und PGM ₁ ⁷ wurde in 7 Familien mit ausreichenden Daten nachgewiesen.

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Established and supported by Stiftung Volkswagenwerk.