Isoelectric focusing of human red cell phosphoglucomutase

Summary A total of 637 individuals from the rural village of Keneba in The Gambia, West Africa, have been typed for red cell PGM using isoelectric focusing (pH 5–7) in polyacrylamide gels. Eight different phenotypes have been detected. The frequency of the four alleles at the PGM₁ locus was found to be PGM ₁ ¹⁺ 0.795, PGM ₁ ^1- 0.053, PGM ₁ ²⁺ 0.133, and PGM ₁ ^2- 0.019. A study of the PGM phenotypes in 89 families confirmed the simple Mendelian codominant inheritance of the four alleles. Comparative population data suggest that red cell PGM typing by isoelectric focusing might prove to be a useful genetic marker in anthropological studies.     

Investigations on the PGM 1 a polymorphism (phosphoglucomutase-EC 2.7.5.1) by isoelectric focusing

Summary The determination of phosphoglucomutase (PGM₁) phenotypes was performed by isoelectric focusing on samples from 1678 unrelated individuals from Hessen, Germany. Ten common phenotypes are considered as gene products of four alleles at the PGM₁ locus with the following frequencies: PGM ₁ ^a1 =0.6305, PGM ₁ ^a2 =0.1844, PGM ₁ ^a3 =0.1320, and PGM ₁ ^a4 =0.0530. Twenty-two different mating types were observed in 113 families with 202 children. The segregation of the phenotypes in the offspring supports the assumed way of autosomal codominant inheritance. The example of a silent allele (PGM ₁ ⁰ ) as well as a rare variant (PGM ₁ ⁷ ) is reported.     

A new hereditary variant of the PGM1 erythrocyte enzyme system determined by isoelectric focusing

Summary A new variant of the PGM _a ¹ erythrocyte enzyme system not identical with the known variants of the system has been detected in the hemolyzed red blood cells of a healthy blood donor by isoelectric focusing. Using this technique the variant is represented by two bands, a strong and slow one more cathodically located than the a3 band and a weak one in the position of the a2 band. Using agarose thinlayer or acetate foil electrophoresis the variant is represented only by a minimal cathodic broadening of the PGM₁ 1 band and therefore it is easily overlooked. Investigation of the propositus' family shows that the variant occurs combined with the common alleles PGM ₁ ^a1 , PGM ₁ ^a2 , and PGM ₁ ^a3 and that it has an autosomal dominant inheritance. Obviously the variant has a very low frequency.     

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     

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.

     

Evidence for two additional common alleles at the PGM1 locus (Phosphoglucomutase-E.C.:2.7.5.1)

Summary Lysates of erythrocytes, leukocytes, lymphocytes, and extracts of sperms were investigated for the PGM₁ isozymes by three techniques: starch gel electrophoresis, high voltage thin-layer agarose gel electrophoresis, and thinlayer isoelectric focusing on polyacrylamide gel. On starch, only the well known common phenotypes 1, 2-1, and 2 were demonstrable. On agarose, different distances of the two main cathodal bands (a, b) among the phenotypes 2-1 were noted. Furthermore, on agarose, some types considered as homozygous on starch gel had a single, sharp banded pattern, while others were broad and blurred. Optimal separation was achieved by isoelectric focusing on polyacrylamide gel. In 291 leukolysates, 10 different phenotypes were identified. These are considered as gene products of 4 different common alleles at the PGM₁ locus as suggested by preliminary family investigations. In a random population from Hessen these four alleles, had the following frequencies: PGM ₁ ^a1 0.6186, PGM ₁ ^a2 0.1718, PGM ₁ ^a3 0.1426, and PGM ₁ ^a4 0.067. The preliminary designation a1, a2, a3 and a4 was chosen as the assumed polymorphism was demonstrated on acrylamide and agarose. The sum of the frequencies PGM ₁ ^a1 and PGM ₁ ^a3 (the gene products of which have apparently the same electrophoretic mobility on starch) is similar to the frequency of the old PGM ₁ ¹ allele (0.757) in Caucasoids, PGM ₁ ^a2 and PGM ₁ ^a4 have a frequency of 0.2388 corresponding with the frequency of the old allele PGM ₁ ² .     

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.     

Purification and Partial Characterization of the Phosphoglucomutase Isozyhes from Human Placenta

We have developed a simple procedure for the purification of phosphoglucomutase (PGM) isozymes from human placenta of healthy women. The technique involves the ammonium sulfate fractionation, ion-exchange and dye-ligand chromatographies. By this method we obtained homogeneous isozyme preparations of the products (“primary” and “secondary”) of the two PGM¹ and PGM² loci. The final specific activities were 1134.6–1441.8 units/mg for PGM¹ forms and 40.2–46.5 units/mg for PGM² forms. On SDS-polyacrylamide gel electrophoresis analysis, the final preparations gave a single protein band of 58,500 and 69,000 Mr for PGM¹ and PGM² isozymes, respectively. These forms have the same kinetic properties, but from the substrate specificity experiments we have found that PGM² forms are more effective for catalyzing the phosphoribomutase and glucose 1,6-bisphosphate synthase reaction than PGM¹ forms. All these properties are shared by the same isozymes previously isolated from human erythrocytes but in this procedure the use of human placenta for the PGM isozymes purification takes advantage of high specific activity of PGM in the extracts of this tissue as well as obtaining highly homogeneous protein suitable for studies at molecular level.     

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.     

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 .     

Population genetic studies of the Aka Pygmies (Central Africa)

Summary Blood samples collected in a single Pygmy tribe, the Aka, living in Bokoka district (Central African Empire) were investigated with respect to the phenotype and gene frequencies of the following 12 enzyme systems: acid phosphatase, adenosine deaminase, adenylate kinase, carbonic anhydrase, esterase D, glucose-6-phosphate dehydrogenase, malate dehydrogenase, phosphoglucomutase 1, phosphoglucomutase 2, phosphogluconate dehydrogenase, superoxide dismutase and serum cholinesterase variants (locus E₁ and E₂). The data obtained in the study of genetic polymorphisms of this isolated and inbred population show a specific pattern with the following characteristics: the very low frequency of PGD^B and p^a alleles; the existence of two rare PGM variants at the PGM₂ locus, typical PGM ₂ ^6Pyg (4.2%) and PGM ₂ ⁹ (0.2%); the high frequency of the p^r allele (10.8%) and CA _II ² (8.22%) and ESD² genes (18.4%). Furthermore, at the G6PD locus four distinct alleles have been found: the negroid GDA-(4%) and GdA+(16%), the common GdB+(79.2%)-,and the rare Gd+^{Ibadan Austin} (0.7%). Cholinesterase typings disclosed the presence of the uncommon E ₁ ^f and E ₁ ^s genes distributed within a single breeding unit. The results are compared with other data previously reported on South African Khoisan and some Negroid populations; the particular genetic background of Pygmies is discussed.Otherwise known as Bi Aka     

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.     

The phosphoglucomutase (PGM1)-groups in the Swiss population

Summary The distribution of the phosphoglucomutase(PGM₁)-groups was studied on blood samples obtained from 2638 Swiss adults. The distribution was found to be in excellent agreement with the Hardy-Weinberg equilibrium. The obtained gene frequencies were similar to those observed in other Caucasian populations (PGM ₁ ¹ =0.7586, PGM ₁ ² =0.2414). In 942 mother/child pairs no theoretical impossible combinations were found. No significant difference was observed between the gene frequencies of men and of women. An unusual phenotype, probably 3-1, was found in blood samples from 3 unrelated adults (1 woman and 2 men). In addition 2 children (a child of the woman and a child of one of the men) were found to have this rare phenotype.

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Zusammenfassung An einem Untersuchungsgut von 2638 Blutproben von schweizerischen Erwachsenen wurde die Verteilung der Phosphoglucomutase(PGM₁)-Gruppen untersucht. Die gefundene Verteilung ist in ausgezeichneter Übereinstimmung mit dem Hardy-Weinberg-Gesetz. Die Frequenzen stimmen mit denen anderer kaukasischer Bevölkerungen überein (PGM ₁ ¹ =0,7586, PGM ₂ ¹ -0,2414). In 942 Mutter/Kind-Paaren wurden keine theoretisch unmögliche Kombination gefunden. Es bestand keine signifikante Differenz zwischen den Genfrequenzen von Frauen und Männern. Bei 3 nichtverwandten Personen (1 Frau, 2 Männer) wurde ein seltener Phänotyp (wahrscheinlich 3-1) beobachtet. Der gleiche Typ wurde bei 2 Kindern gefunden (eines war das Kind der Frau, das andere dasjenige von einem der Männer).

     

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.     

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).     

Phosphoglucomutase (EC 2.7.5.1.) and adenylate kinase (EC 2.7.4.3.) typings in Koreans and Irish

Summary PGM₁ and AK phenotypes were determined in samples from Korea and Ireland. the frequencies of PGM ₁ ¹ genes amount to 0.916 in Koreans and 0.864 in Irish. AK¹ frequencies come to 0.933 in Koreans and 0.873 in Irish.Supported by the Deutsche Forschungsgemeinschaft.     

The distribution of serum protein and enzyme groups among the Batak of Samosir Island (Sumatra,Indonesia)

Summary 188 blood samples from Batak of Samosir Island (Sumatra, Indonesia) have been studied for electrophoretic variants of haemoglobin, 14 red cell enzyme and 5 serum protein systems. The acid phosphatase, 6 PGD, PGM₁ and ADA enzyme systems are polymorphic, and a single AK 2-1 person was detected. Polymorphism is present in the haptoglobin, transferrin and protease inhibitor systems. Two variant alleles, Tf ^Dchi and Tf ^B are present in the transferrin system, but the B variant has not been identified. Similarly, 3 persons with caeruloplasmin variants were found, but also these variants have not been identified. No abnormal haemoglobins were detected. All other systems revealed the presence of only normal phenotypes.

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Zusammenfassung 188 Blutproben des Batak-Stammes von Samosir Island (Sumatra, Indonesien) wurden auf elektrophoretische Varianten des Hämoglobins in 14 Erythrocyten-enzym-und 5 Serumprotein-Systemen untersucht. Die Saure Phosphatase, 6GPD, PGM₁ und ADA-Systeme sind polymorph, und eine einzige AK 2-1-Person wurde gefunden. In den Haptoglobin-, Transferrin- und Pi-Systemen treten Polymorphismen auf. Zwei abweichende Allele, Tf ^Dchi und Tf ^B, sind im Transferrin-System zu finden, aber die B-Variante ist nicht bestimmt worden. Ebenso wurden 3 Personen mit Ceruloplasmin-Varianten gefunden, doch auch diese Varianten wurden nicht identifiziert. Keine abnormen Hämoglobine wurden gefunden. Alle anderen Systeme wiesen nur normale Phenotypen auf.

     

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.     

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.