Power of assaying inbreeding through sampling of phenotypes and mating types

Summary Several enzyme polymorphisms and hemoglobin variants were typed in a sample of n=219 non-related Greek blood-donors. The following gene frequencies were observed: p^a=0.201, p^b=0.701, p^c=0.098; PGD^A=0.985, PGD^c=0.015; AK¹=0.942, AK²=0.058; Hb^A=0.988, Hb^S=0.012. No polymorphic variation was seen in LDH, s-MDH, PHI, or SOD. The population genetical aspects of these results are discussed.

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Zusammenfassung An einer Stichprobe von n=219 erwachsenen griechischen Blutspendern wurden verschiedene Enzympolymorphismen und Hämoglobinvarianten untersucht. Folgende Genfrequenzen konnten beobachtet werden: p^a=0,201, p^b=0,701, p^c=0,098; PGD^A=0,985, PGD^c=0,015; AK¹=0,0942, AK²=0,058; Hb^A=0,988, Hb^S=0,012. Keine polymorphe Variation wurde bei LDH, s-MDH, PHI und SOD gefunden. Die populationsgenetischen Aspekte dieser Befunde werden diskutiert.

     

Adenylate kinase polymorphism in populations in Finland (Swedes,Finns, Lapps), in Maris,and in Greenland Eskimos

Summary Starch-gel electrophoresis for adenylate kinase (AK) was performed on 2519 haemolysates from 6 population samples of unrelated males in Finland, 4 Finnish Lapp populations, the Maris (Cheremisses) in the USSR, and an Eskimo population in NW Greenland. Between the Finland Swedes and Finns no significant difference in AK polymorphism was observed and the allele frequency estimates of AK were comparable with those found in other Europeans. The indigenous pure Skolt Lapps showed absence of the AK² gene, which was also extremely low in the Fisher and Mountain Lapps. All the Lapp populations so far studied show an extremely low frequency of the AK² gene. The rarity of AK² in Lapps may offer a better approach to the estimation of intermixture than certain other genes which vary in frequency in different Lapp populations. Among the Maris AK² frequencies are lower (0.017) than among other Europeans. The AK² was also very low (0.016) in the Greenland Eskimo population on Augpilagtok Island.The results obtained for the AK phenotypes in 149 Lapp families and in 84 mothers and their children are in agreement with the hypothesis that AK¹ and AK² are alleles at one and the same autosomal locus. The present family and mother-child studies add further evidence for the acceptance of the AK system as a valuable tool in cases of disputed paternity.Supported by the Finnish National Research Council for Medical Sciences, the Finnish Academy of Science and Letters, the Wenner-Gren Foundation for Anthropological Research and the Deutsche Forschungsgemeinschaft. Preparations were partly obtained as a gift from Boehringer GmbH, Mannheim, Germany.     

Red cell enzyme polymorphisms in Bulgaria

Summary 7 human red cell enzyme polymophisms have been typed on a sample of n=138 unrelated adults from Bulgaria, which revealed the following gene frequencies: ADA¹=0.8623. ADA²=0.1376; AK¹=0.9637, AK²=0.0362; 6-PGD^A=0.9891, 6-PGD^C=0.0108; PGM ₁ ¹ =0.8346, PGM ₁ ² =0.1653; P^A=0.1596, P^B=0.7983, P^C=0.0420. In the LDH-system one B-subunit variant was found, whilst no Peptidase A or B variant could be observed. The anthropological significance of these findings is discussed.

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Zusammenfassung An einer Stichprobe von n=138 nichtverwandten erwachsenen Bulgaren wurden 7 erythrocytäre Enzympolymorphismen untersucht. Dabei ergaben sich die folgenden Frequenzen: ADA¹=0,8623, ADA²=0,1376; AK¹=0,9637, AK²=0,0362; 6-PGD^A=0,9891, 6-PGD^C=0,0108; PGM ₁ ¹ =0,8346, PGM ₁ ² =0,1653; P^A=0,1596, P^B=0,7983, P^C=0,0420. Im LDH-System wurde eine B subunit-Variante gefunden, während keine Peptidase A- oder B-Variante beobachtet werden konnte. Die anthropologische Bedeutung dieser Befunde wird diskutiert.

     

Crystal structure of a trimeric archaeal adenylate kinase from the mesophile Methanococcus maripaludis with an unusually broad functional range and thermal stability

The structure of the trimeric adenylate kinase from the Archaebacteria Methanococcus mariplaludis (AK_MAR) has been solved to 2.5‐Å resolution and the temperature dependent stability and kinetics of the enzyme measured. The K_M and V_max of AK_MAR exhibit only modest temperature dependence from 30°–60°C. Although M. mariplaludis is a mesophile with a maximum growth temperature of 43°C, AK_MAR has a very broad functional range and stability (T_m = 74.0°C) that are more consistent with a thermophilic enzyme with high thermostability and exceptional activity over a wide range of temperatures, suggesting that this microbe may have only recently invaded a mesophilic niche and has yet to fully adapt. A comparison of the Local Structural Entropy (LSE) for AK_MAR to the related adenylate kinases from the mesophile Methanococcus voltae and thermophile Methanococcus thermolithotrophicus show that changes in LSE are able to fully account for the intermediate stability of AK_MAR and highlights a general mechanism for protein adaptation in this class of enzymes. Proteins 2010. © 2009 Wiley‐Liss, Inc.     

Purine excretion by mammalian cells deficient in adenosine kinase

An adenosine kinaseless (AK^?) mutant of the mouse fibroblast line 3T6 has been obtained in cell culture by evolution of resistance to 6-thio-methylpurine ribonucleoside and tubercidin. The mutant excretes purines (xanthine and hypoxanthine) into the culture medium. Human or mouse cells lacking hypoxanthine-guanine phosphoribosyl transferase (HPT^?) excrete increased amounts of purines, but a human cell mutant lacking both HPT and AK excretes considerably more hypoxanthine. The difference in hypoxanthine excretion between the HPT^? mutant and the HPT^? AK^? mutant originates from the adenosine normally reutilized through the activity of adenosine kinase. The activity of adenosine kinase is essential to retard the adenosine cycle and to prevent cellular loss of purines.     

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.     

Calculation of the maximum amount of gene admixture in a hybrid population

Evidence is presented to show that “Caucasian” genes (B, K, Lu^a, r, AK², P^c, and Gm^3,5,11) in hybrid North American Indian populations follow a Poisson distribution. A method of determining the maximum amount of admixture, given an observed count of Caucasian genes, is developed. Establishment of the upper limit of admixture is suggested as the preferred estimate of gene flow in situations where absence of specific genes at particular loci precludes the calculation of a mean admixture estimate.     

Zur Populationsgenetik der Adenylatkinase: Genhäufigkeit in einer südwestdeutschen Stichprobe

Summary Adenylate kinase phenotypes were determined in 407 unrelated persons from Southwestern germany. The frequencies were estimated to be AK¹=0.969, AK²=0.031.

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Direktor: Prof. Dr. med. Dr. H. Baitsch

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Release of adenosine by C1300 neuroblastoma cells in tissue culture

Previous work in our laboratory led us to postulate that N2a cells release adenosine into growth medium, where it acts at the extracellular adenosine receptors to modulate the sensitivity of the cells to the cyclic AMP-elevating effect of adenosine [Green, RD, J Pharmacol Exp Ther 201:610, 1977]. We have now devised a high-performance liquid chromatographic (HPLC) procedure capable of quantitating the concentrations of adenosine in cells and tissue culture media. Growth media of N2a cells and a variant of N2a cells deficient in hypoxanthine-guanine phosphoribosyltransferase (HGPRT^?) contain 10–20 nM adenosine, while that of a variant deficient in adenosine kinase (AK^?) is elevated severalfold. It appears that the concentration of adenosine in growth media is determined by both the rate at which it is released by cells into the medium and the rate at which it is metabolized by adenosine deaminase present in the serum in the growth medium. Both N2a and AK^? cells release considerable amounts of adenosine into serum-free medium (SFM) over a short period. Adenosine release is greater from AK^? cells and is accelerated by erythro-9-(2-hydroxy-3-nonyl)-adenine (EHNA), a potent adenosine deaminase inhibitor. This accelerated release is retarded by dipyridamole and homocysteine. Surprisingly, dipyridamole and 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (Ro 20 1724), a potent phosphodiesterase inhibitor, stimulate basal adenosine release from N2a but not from AK^? cells. It remains to be determined if this is due to an effect of these compounds on adenosine kinase. These results give further support for the hypothesis that adenosine in growth medium modulates the sensitivity of the cells to the cyclic AMP-elevating affect of adenosine, and furthermore they suggest that adenosine in growth media may tonically stimulate adenylate cyclase and affect processes controlled by the cyclic AMP:cyclic AMP-dependent protein kinase system.     

AK,ADA and 6-PGD polymorphisms in Cosenza province (Calabria,Italy)

We report the allele frequencies of polymorphic red cells adenilatechinase (AK), adenosindeaminase (ADA) and 6-phosphogluconate dehydrogenase (6-PGD) enzymes in a population living in the province of Cosenza (Calabria, Southern Italy). AK^*1 (0.957), ADA^*1 (0.942) and 6-PGD^*A (0.965) frequencies are compared with samples obtained from other italian regions.     

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.     

Studies on blood groups and other genetic markers in forest Nentzi: Variation among the subpopulations

Summary A Samoyed-speaking group of fishers, hunters, and deer breeders numbering 1500 in total has been investigated. Seven territorial subgroups were examined with respect to 15 genetic systems. The presence of A₂, cde, C^wDe, Kp^a, AK², p^c, Gm^fb, and Gm^fnb genes or haplotypes in low or moderate frequencies was observed. An unexpected finding was a deleted Gm(-;n;gb) phenotype in three siblings.Significant local genetic heterogeneity was observed with regard to AB0, Rh, Tf, PGM₁, 6-PGD, and Gm systems. The summed genic heterogeneity was found to be highly significant ( ₁₃₂ ² =663.70, P< 0.001). Mean F_st was equal to 0.0228, reflecting variation among subpopulations dispersed in the forest-tundra area and living under conditions of partial isolation.     

Anthropological studies among Libyans. Erythrocyte genetic factors,serum haptoglobin phenotypes and anthropometry

Anthropological studies were done on 1276 Libyans from the Mediterranean cities of Tripoli and Benghazi, and from Sabha southward in The Sahara. The incidences of hemoglobin (Hb)-S and glucose-6-phosphate dehydrogenase (G-6-PD) deficiency were low in the coastal areas and significantly high in Sabha. Hb-C occurred sporadically in Tripoli and Sabha, and was absent from Benghazi in the east. One case of Hb-J Benghazi was noted. There were no significant differences in the ABO blood group and Rh₀ (D) type distributions in the three localities. G-6-PD gene Gd^A frequency was significantly high in Sabha. The lowest value of 6-phosphogluconate dehydrogenase (6-PGD) gene PGD^A frequency and highest value of the gene PGD^C were in Sabha. Adenylate kinase (AK) gene AK² was only detectable in Tripoli. Acid phosphatase (AP) gene P^a frequency in Sabha was more than twice that in Tripoli and Benghazi, while P^c was distinctly lower in Sabha than in the northern cities. Haptoglobin gene Hp¹ frequency was almost identical in all areas. Anthropometric measurements revealed overall homogeneity of the three samples, closer similarity in the coastal region to adjacent North African populations, and Negroid influence in the Saharan Libyans. Anthropometry substantiated findings from blood markers.     

The adnylate kinase (AK)-groups in the Swiss population

Summary The distribution of the adenylate kinase groups was studied on blood samples obtained from 2270 Swiss individuals. 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 (AK¹=0.957, AK²=0.043). In 335 mother/childpairs no theoretically impossible combinations were found. No significant difference was observed between the gene-frequencies of men and women. Among the blood samples investigated an unusual phenotype was found in one mother and her child.     

The biological role of octopine in the squid,Loligo vulgaris (Lamarck)

Summary The enzymatic activities of glyceraldehyde-3-phosphate dehydrogenase, octopine dehydrogenase and lactate dehydrogenase were determined fromLoligo vulgaris. Octopine dehydrogenase displays the highest activity yet recorded for this enzyme, exceeding glyceraldehyde-3-phosphate dehydrogenase sixfold and lactate dehydrogenase 365-fold (Table 1).During jet propulsion swimming octopine accumulates instead of lactate (Table 2), while phosphoarginine, the phosphagen of the squid, is depleted (Table 3).The formation of octopine is discussed in relation to anaerobic metabolism which might occur during burst activity in cephalopods.The following abbreviations are used AK arginine kinase (2.7.3.3) - GAPDH glyceraldehyde-3-phosphate dehydrogenase (1.2.1.12) - LDH L-lactate - NAD oxidoreductase (1.1.1.27) - ODH octopine - NAD oxidoreductase (1.5.1.11) - DTT dithiothreitol - dw dry weight (about 20% of the fresh weight) This investigation was generously supported by The Deutsche Forschungsgemeinschaft grant No.: (Ze 40/13)     

Daten zur Populationsgenetik der 6-Phosphogluconat-Dehydrogenase

Summary Four Malaysian racial groups were typed for red cell adenylate kinase: 324 Malays, 300 Chinese, 256 Indians, and 483 West Malaysian Aborigines. The AK² gene frequencies found were 0.015, 0.0, 0.086, and 0.013, respectively. All 244 Malays, 170 Chinese, 153 Indians, and 132 West Malaysian Aborigines examined had the common cytoplasmic malate dehydrogenase phenotype.

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Zusammenfassung Vier Rassengruppen aus Malaysia wurden auf Adenylatkinase-Varianten hin untersucht: 324 Malayen, 300 Chinesen, 256 Inder und 483 Eingeborene von West-Malaysia. Die Genhäufigkeiten des Allels AK² waren: 0,015, 0,0, 0,086 und 0,013. Alle 244 Malayen, 170 Chinesen, 153 Inder und 132 west-malaysischen Eingeborenen, die daraufhin untersucht werden konnten, hatten den häufigen Phänotyp der cytoplasmatischen Malat-Dehydrogenase.

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This work was supported by the University of Maryland and University of California International Centers for Medical Research and Training with research grants AI-10049-12, AI-10051, and HE 10486, all from the National Institutes of Health, U.S. Public Health Service.     

Red cell enzyme polymorphisms in Punjabis in North India

Seven red cell isoenzyme systems were investigated on a sample of 140 Punjabis from Hoshiarpur and Chandigarh, shown to be representative by comparison of their blood group frequencies with other samples from the area. Phenotype and gene frequencies are given for adenosine deaminase, adenylate kinase, acid phosphatase, 6-phosphogluconate dehydrogenase, phosphoglucomutase locus 1 and 2, lactate dehydrogenase and phosphohexose isomerase. The high frequencies of the ADA² and AK² genes in Indian samples and the presence of the rare variant 3-1 of phosphohexose isomerase are confirmed.     

No association of red cell adenylate kinase phenotypes with affective disorders

Summary Red cell adenylate kinase (AK) phenotypes were studied in 195 patients with affective disorders (41 with the bipolar and 81 with the unipolar form of the disease) and 418 controls. No significant differences were found between patients and controls and between patients with different types of affective disorders. Thus the previous observation by Rundle et al. (1977) showing an increased frequency of the AK² allele in the unipolar group was not confirmed.     

Characterization of aspartate kinase and homoserine dehydrogenase from <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis> IWJ001 and systematic investigation of <Emphasis Type="SmallCaps">l</Emphasis>-isoleucine biosynthesis

Previously we have characterized a threonine dehydratase mutant TD^F383V (encoded by ilvA1) and an acetohydroxy acid synthase mutant AHAS^{P176S, D426E, L575W} (encoded by ilvBN1) in Corynebacterium glutamicum IWJ001, one of the best l-isoleucine producing strains. Here, we further characterized an aspartate kinase mutant AK^A279T (encoded by lysC1) and a homoserine dehydrogenase mutant HD^G378S (encoded by hom1) in IWJ001, and analyzed the consequences of all these mutant enzymes on amino acids production in the wild type background. In vitro enzyme tests confirmed that AK^A279T is completely resistant to feed-back inhibition by l-threonine and l-lysine, and that HD^G378S is partially resistant to l-threonine with the half maximal inhibitory concentration between 12 and 14 mM. In C. glutamicum ATCC13869, expressing lysC1 alone led to exclusive l-lysine accumulation, co-expressing hom1 and thrB1 with lysC1 shifted partial carbon flux from l-lysine (decreased by 50.1 %) to l-threonine (4.85 g/L) with minor l-isoleucine and no l-homoserine accumulation, further co-expressing ilvA1 completely depleted l-threonine and strongly shifted carbon flux from l-lysine (decreased by 83.0 %) to l-isoleucine (3.53 g/L). The results demonstrated the strongly feed-back resistant TD^F383V might be the main driving force for l-isoleucine over-synthesis in this case, and the partially feed-back resistant HD^G378S might prevent the accumulation of toxic intermediates. Information exploited from such mutation-bred production strain would be useful for metabolic engineering.