Hum. Herod. 26: 161-166 (1976)
Polymorphism of Human Red Cell Adenosine Deaminase, Esterase D, Glutamate Pyruvate Transaminase and Galactose1-Phosphate-Uridvltransferase in the Swiss Population R .P flugshaupt , R.SciiERzand R. Butler Central Laboratory, Blood Transfusion Service, Swiss Red Cross, Berne
Key H/onk. Adenosine deaminase • Esterase D • Glutamate pyruvate transaminase • Galactose-l-phosphate-Uridyltransferase • Gene frequencies • Swiss population Abstract. The distribution of the phenotypes of the red cell enzymes adenosine de aminase (EC 3.5.4.4), esterase D, glutamate pyruvate transaminase (EC 2.6.1.2) and galactose-l-phosphate-uridyltransferase (EC 2.7.7.12). were studied on a large number of Sw iss individuals. The phenotypes of all four enzymes were found to be distributed accord ing to the Hardy-Weinberg law , although lescls of statistical significance of the differences were not of the same order for all four enzymes. Gene frequencies were calculated and found to be in good agreement with data from the literature concerning the Caucasian populations. Among numerous mother-child pairs, no phenotype constellations that were in contradiction to the genetical hypothesis were observed. Phenotypes were distributed identically in men and women.
The enzyme adenosine deaminase (ADA; EC 3.5.4.4) acts as an aminohydrolase catalyzing the deamination of adenosine into inosine. It was found in different tissues and in red blood cells. The enzyme is known to exist in multiple molecular forms. S pencer et at. [22] studied its polymorphism in erythrocytes for the lirst time by starch-gel electrophoresis. They found three common phenotypes called ADA 1, ADA 2-1, and ADA 2. Since then, several additional rare phenotypes have been observed, which were attributed to the alleles ADAS, ADA4, ADA’’, ADAB [19]. In addition, a ‘silent’ allele ADA" has been described [6, 7], which appears to be associated with combined immunodeficiency. A polymorphism of a red cell esterase (EsD) has been detected by Hop KINSON et al. [II]. According to its substrate specificity to umbellifcryl acetate
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Introduction
162
P l L U G S IIA lJl“l / SC’H F R / BU TLER
or butyrate it could be distinguished from the formerly known esterases A, B, C. It was therefore referred to as esterase D. Until now three common phenotypes have been described, namely EsD I, EsD 2-1 and EsD 2. Yet another much rarer phenotype EsD 3 - 1 has been reported by Bend er and F r a n k [I], indicating the existence of a third allele EsD'1. Glutamate pyruvate transaminase (GPT; EC 2.6.1.2), synonyme alanine aminotransferase, catalyzes the conversion of L-alanine and u-ketoglutarate into L-glutamate and pyruvate. The enzyme exists in a mitochondrial or granular form and in the cytoplasmatic or soluble form. Soluble GPT of human red cells show a polymorphism when separated in starch-gel electro phoresis [5], Three common phenotypes called GPT I. GPT 2-1 and GPT 2 are distinguishable. A number of rare alleles, GPT3, GPT'. GPT5, GPT*’. GPT7, and the existence of a ‘silent’ gene GPT" have been reported [9, 16, 23], The enzymatic activities differ according to the different phenotypes [24], Galactose-I-phosphate-uridyltransferase (Gt; EC 2.7.7.12) catalyzes the conversion of galactose-l-phosphate into glucose-l-phosphate. The complete absence of activity of this enzyme leads to the severe inborn error of meta bolism referred to as galactosemia. A silent allele, Gt°, at the Gt locus was postulated. Galactosemic patients are homozygous in this respect (Gt°/Gt°). Family studies revealed that heterozygotes had approximately one half of the enzyme activity. B eutler et al. [3] discovered a variant of the enzyme which had yet a different activity. They called it Duarte variant. Further studies proved that this variant was due to an allelic gene, G tDuarle, or Gt2, as it is called now. In 1973, an additional gene product, the Los Angeles variant, was discovered by No el al. [14]. The different phenotypes of the enzyme can be separated by electrophoresis on starch gel or agarose gel, respectively. An autosomal codominant mode of inheritance is known for all four enzyme systems. Material and Methods
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Blood samples from Swiss blood donors and from unrelated people engaged in cases of disputed paternity were typed for the four enzymes by starch-gel electrophoresis or agarose-gel electrophoresis. Red cells were washed 3 times with saline and lyzed by freezing and thawing. Hcmolyzates were submitted to electrophoresis. ADA and EsD phenotypes were separated in the same starch gel together with adenyl ate kinase (AK). The method of separation has been described elsewhere [18], Under these conditions, ADA and EsD migrate towards the anode, while AK is migratingcathodically.
163
Swiss Gene Frequencies of ADA, EsD. GPT. Gt Table I. Distribution of ADA, EsD, GPT and Gt phenotypes in Swiss individuals
ADA ADA ADA ADA ADA v EsD EsD EsD EsD
GPT GPT GPT GPT GPT GPT r Gt Gt Gt Gt Gt V
n = 6,604 1 2-1 2 4-1
n = 744 1 2-1 2
n = 1,894 1 2 1 2 3-1 3 2
n = 1,098 1 2-1 2 R-I
11
Observed %
Expected n
%
5,828 763 12 1 6.604
88.2 11.6 0.2
Polymorphism of Human Red Cell Adenosine Deaminase, Esterase D, Glutamate Pyruvate Transaminase and Galactose1-Phosphate-Uridvltransferase in the Swiss Population R .P flugshaupt , R.SciiERzand R. Butler Central Laboratory, Blood Transfusion Service, Swiss Red Cross, Berne
Key H/onk. Adenosine deaminase • Esterase D • Glutamate pyruvate transaminase • Galactose-l-phosphate-Uridyltransferase • Gene frequencies • Swiss population Abstract. The distribution of the phenotypes of the red cell enzymes adenosine de aminase (EC 3.5.4.4), esterase D, glutamate pyruvate transaminase (EC 2.6.1.2) and galactose-l-phosphate-uridyltransferase (EC 2.7.7.12). were studied on a large number of Sw iss individuals. The phenotypes of all four enzymes were found to be distributed accord ing to the Hardy-Weinberg law , although lescls of statistical significance of the differences were not of the same order for all four enzymes. Gene frequencies were calculated and found to be in good agreement with data from the literature concerning the Caucasian populations. Among numerous mother-child pairs, no phenotype constellations that were in contradiction to the genetical hypothesis were observed. Phenotypes were distributed identically in men and women.
The enzyme adenosine deaminase (ADA; EC 3.5.4.4) acts as an aminohydrolase catalyzing the deamination of adenosine into inosine. It was found in different tissues and in red blood cells. The enzyme is known to exist in multiple molecular forms. S pencer et at. [22] studied its polymorphism in erythrocytes for the lirst time by starch-gel electrophoresis. They found three common phenotypes called ADA 1, ADA 2-1, and ADA 2. Since then, several additional rare phenotypes have been observed, which were attributed to the alleles ADAS, ADA4, ADA’’, ADAB [19]. In addition, a ‘silent’ allele ADA" has been described [6, 7], which appears to be associated with combined immunodeficiency. A polymorphism of a red cell esterase (EsD) has been detected by Hop KINSON et al. [II]. According to its substrate specificity to umbellifcryl acetate
Downloaded by: King's College London 137.73.144.138 - 1/16/2019 2:34:17 AM
Introduction
162
P l L U G S IIA lJl“l / SC’H F R / BU TLER
or butyrate it could be distinguished from the formerly known esterases A, B, C. It was therefore referred to as esterase D. Until now three common phenotypes have been described, namely EsD I, EsD 2-1 and EsD 2. Yet another much rarer phenotype EsD 3 - 1 has been reported by Bend er and F r a n k [I], indicating the existence of a third allele EsD'1. Glutamate pyruvate transaminase (GPT; EC 2.6.1.2), synonyme alanine aminotransferase, catalyzes the conversion of L-alanine and u-ketoglutarate into L-glutamate and pyruvate. The enzyme exists in a mitochondrial or granular form and in the cytoplasmatic or soluble form. Soluble GPT of human red cells show a polymorphism when separated in starch-gel electro phoresis [5], Three common phenotypes called GPT I. GPT 2-1 and GPT 2 are distinguishable. A number of rare alleles, GPT3, GPT'. GPT5, GPT*’. GPT7, and the existence of a ‘silent’ gene GPT" have been reported [9, 16, 23], The enzymatic activities differ according to the different phenotypes [24], Galactose-I-phosphate-uridyltransferase (Gt; EC 2.7.7.12) catalyzes the conversion of galactose-l-phosphate into glucose-l-phosphate. The complete absence of activity of this enzyme leads to the severe inborn error of meta bolism referred to as galactosemia. A silent allele, Gt°, at the Gt locus was postulated. Galactosemic patients are homozygous in this respect (Gt°/Gt°). Family studies revealed that heterozygotes had approximately one half of the enzyme activity. B eutler et al. [3] discovered a variant of the enzyme which had yet a different activity. They called it Duarte variant. Further studies proved that this variant was due to an allelic gene, G tDuarle, or Gt2, as it is called now. In 1973, an additional gene product, the Los Angeles variant, was discovered by No el al. [14]. The different phenotypes of the enzyme can be separated by electrophoresis on starch gel or agarose gel, respectively. An autosomal codominant mode of inheritance is known for all four enzyme systems. Material and Methods
Downloaded by: King's College London 137.73.144.138 - 1/16/2019 2:34:17 AM
Blood samples from Swiss blood donors and from unrelated people engaged in cases of disputed paternity were typed for the four enzymes by starch-gel electrophoresis or agarose-gel electrophoresis. Red cells were washed 3 times with saline and lyzed by freezing and thawing. Hcmolyzates were submitted to electrophoresis. ADA and EsD phenotypes were separated in the same starch gel together with adenyl ate kinase (AK). The method of separation has been described elsewhere [18], Under these conditions, ADA and EsD migrate towards the anode, while AK is migratingcathodically.
163
Swiss Gene Frequencies of ADA, EsD. GPT. Gt Table I. Distribution of ADA, EsD, GPT and Gt phenotypes in Swiss individuals
ADA ADA ADA ADA ADA v EsD EsD EsD EsD
GPT GPT GPT GPT GPT GPT r Gt Gt Gt Gt Gt V
n = 6,604 1 2-1 2 4-1
n = 744 1 2-1 2
n = 1,894 1 2 1 2 3-1 3 2
n = 1,098 1 2-1 2 R-I
11
Observed %
Expected n
%
5,828 763 12 1 6.604
88.2 11.6 0.2
