Genetic variation at the alcohol dehydrogenase locus inDrosophila melanogaster: A third ubiquitous allele

Summary A 3rd allele at theAdh locus,Adh ^FCh.D., has been found at polymorphic frequencies in natural populations ofD. melanogaster. The ADH-FChD enzyme has properties distinct from those of the 2 more common forms of ADH. TheAdh polymorphism should now be analyzed as a triallelic system.     

Guaiacol-O-methyltransferase: A mammalian enzyme capable of forming di-O-methyl catecholamine derivatives

Résumé On a extrait et characterisé une enzyme capable de transférer aux catécholamines 4-O-méthyles le group méthyle provenant de l'S-adénosylméthionine et produire des substances diméthoxyques. Cette enzyme est active dans différents tissus de Mammifères.

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This work was supported by Public Health Service grants Nos. MH-08618 and MH-07961 and a Research Scientist's Award No. MH-14020, to A.J.F.     

Unique evolution of Bivalvia arginine kinases

The clams Pseudocardium, Solen, Corbicula and Ensis possess a unique form of arginine kinase (AK) with a molecular mass of 80 kDa and an unusual two-domain structure, a result of gene duplication and subsequent fusion. These AKs also lack two functionally important amino acid residues, Asp⁶² and Arg¹⁹³, which are strictly conserved in other 40-kDa AKs and are assumed to be key residues for stabilizing the substrate-bound structure. However, these AKs show higher enzyme activity. The cDNA-derived amino acid sequences of 40-kDa AKs from the blood clam Scapharca broughtonii and the oyster Crassostrea gigas were determined. While Asp⁶² and Arg¹⁹³ are conserved in Scapharca AK, these two key residues are replaced by Asn and Lys, respectively, in Crassostrea AK. The native enzyme from Crassostrea and both of the recombinant enzymes show an enzyme activity similar to that of two-domain clam AKs and at least twofold higher than that of other molluskan AKs. Although the replacement of Asp⁶² or Arg¹⁹³ by Gly in normal AK causes a considerable decrease in V_max (6–15% of wild-type enzyme) and a two- to threefold increase in K_m for arginine, the same replacement in Scapharca AK had no pronounced effect on enzyme activity. Together with the observation that bivalve AKs are phylogenetically distinct from other molluskan AKs, these results suggest that bivalve AKs have undergone a unique molecular evolution; the characteristic stabilizing function of residues 62 and 193 has been lost and, consequently, the enzyme shows higher activity than normal.Received 14 October 2003; accepted 1 November 2003     

Laccase production byPolyporus sanguineus under different nutritional and environmental conditions

Summary Laccase production was higher in malt extract medium than in lignin, andPolyporus sanguineus appears to be better thanPolyporus versicolor andTrametes hirsuta (syn.Polyporus hirsutus) for enzyme production. Phenolic compounds, of which resorcinol was the most active, induced enzyme production; while sugars repressed it. A temperature of 37°C, pH 3 and indulin AT at a concentration of 0.2% gave the best enzyme yield.A part of Ph. D. thesis of D.S. Arora.Acknowledgments. We thank Westvaco Chemical Division, USA, and Dr P.S. Rehill, Forest Research Institute, Dehradun, India for providing lignin samples and the fungal strain, respectively. D.S. Arora further thanks University Grants Commission, New Delhi for financial assistance.     

Chromate reduction inStreptomyces

Summary Streptomyces species 3M grew in peptone yeast extract medium with 1000 g/ml K₂Cr₂O₇. Incubation of the chromate with different cell fractions in the presence of NADH and NADPH resulted in a decrease of Cr⁶⁺ in the reaction mixture. The level of Cr⁶⁺ was reduced by 82.7% by a particulate cell fraction obtained by centrifugation at 105,000×g for 1 h, in the presence of NADH. The reducing enzyme was associated with this cell fraction. The enzyme was constitutive and reduced Cr⁶⁺ to Cr³⁺.     

Repressible alkaline phosphotase inAspergillus niger

Summary ALP fromA. niger is a) P_i repressible enzyme; b) stimulated by addition of Zn⁺⁺ to the growth medium, and c) that EDTA inhibits the enzyme reversibly, which could be restored by addition of Zn⁺⁺ and perhaps Mg⁺⁺. This property is in contrast to the enzyme fromN. crassa, which is independent of any metal requirement.     

Reactivation by glycerol and ethylene glycol of inactivatedδ-aminolevulinic acid synthetase ofRhodopseudomonas spheroides

Summary Reactivation effects by glycerol and ethylene glycol of inactivated ALA synthetase ofR. spheroides were observed. Accompanying the reactivation of the inactivated enzyme, K _m value for PLP decreased to levels similar to those of the freshly prepared enzyme.     

Urinary excretion of renal brush border membrane enzymes in leprosy patientseffect of multidrug therapy

Renal function at the brush border membrane level has been studied using characteristic enzymes, such as alkaline phosphatase, leucine-aminopeptidase and gamma-glutamyl transpeptidase. Urinary enzyme studies were performed using leprosy patients, classified on the basis of bacteriological index (BI>3; n=20,BI<3; n=12, BI-ve; n=10) and compared with control subjects (n=10). The role of enzymuria in monitoring WHO-recommended multidrug therapy (MDT) has been evaluated in these patients. A significant increase in the enzyme activities (p<0.01), as well as significant (p<0.01) proteinurea in 24-hour urine samples of both the smear positive groups (BI>3,BI<3) prior to therapy compared to control subjects, indicates proximal tubular functional impairment at brush border membrane level. In the smear negative (BI-ve) grou, no significant difference was observed in enzyme activities as compared with the control group. In a follow-up study (BI>3; n=13, BI<3; n=4) the activities of all the enzymes decreased significantly in all the groups when compared to a corresponding untreated group. The follow-up study was not carried out on the smear negative group. The surprising finding was the differential behaviour of r-glutamyl transpeptidase, whose activity increased significantly (p<0.01) even after therapy inBI>3 group when compared with untreated patients. However in a detailed work-up including hepatic and renal function tests, the serum biochemistry was found to be normal both before and after therapy. Urinary excretion of brush border enzymes seems to be related to bacterial load, and their potential in studying the effect of MDT remains unclear.     

Effect of benzimidazole on nicotinamide adenine dinucleotide phosphate phosphomonosterase activity in wheat leaves

Summary Nicotinamide adenine dinucleotide phosphate phosphomonoesterase was isolated and partially purified from wheat (Triticum aestivum L. var. Selkirk) leaves. The enzyme hadK _NADP value of 1.4×10^–4 M and a pH optimum of 5.9.In vitro activity of this enzyme was unaffected by precursors of NAD (nicotinamide and nicotinic acid) or cytokinis (kinetin and benzimidazole). However, when detached wheat leaves were treated with solutions of these compounds, the precursors lowered the specific activity while the cytokinins enhanced the activity. It is suggested that spatial separation and compartmentation of the enzyme and its substrate NADP account for the similar effect of benzimidazole on both.This work was supported by a grant No. A2698 from the National Research Council, Canada.     

Beeinflussung der ATPase aminspeichernder Granula von Nebennierenmark und Milznerven durch Thallium

Summary ATPases of the amine storing granules from bovine adrenal medulla and splenic nerves are inhibited by Tl⁺⁺⁺ 3×10^–5 and 5×10^–6 M, respectively. Tl⁺ up to 10^–3 M is ineffective. By T1⁺⁺⁺ in concentrations of 10^–4 M or more, proteins are precipitated, so that the enzyme inhibiton by these concentrations is unspecific. If T1⁺ is oxidized to Tl⁺⁺⁺ in the organism, the inhibition of granular ATPase may be responsible for the alterations of the catecholamine metabolism observed in thallium intoxication.     

The prolyl oligopeptidase family

A group of serine peptidases, the prolyl oligopeptidase family, cannot hydrolyze peptides containing more than about 30 residues. This group is unrelated to the classical trypsin and subtilisin families, and includes dipeptidyl peptidase IV, acylaminoacyl peptidase and oligopeptidase B, in addition to the prototype prolyl oligopeptidase. The recent crystal structure determination of prolyl oligopeptidase (80 kDa) has shown that the enzyme contains a peptidase domain with an α/β hydrolase fold, and its catalytic triad is covered by the central tunnel of an unusual seven-bladed β-propeller. This domain operates as a gating filter, excluding large, structured peptides from the active site. The binding mode of substrates and the catalytic mechanism differ from that of the classical serine peptidases in several features. The members of the family are important targets of drug design. Prolyl oligopeptidase is involved in amnesia, depression and blood pressure control, dipeptidyl peptidase IV in type 2 diabetes and oligopeptidase B in trypanosomiasis. Received 8 August 2001; received after revision 19 September 2001; accepted 21 September 2001     

Endocytosis and inositol hexakisphosphate levels inras transformants ofDictyostelium discoideum amoebae

Summary Fluid-phase pinocytosis kinetics and lysosomal enzyme secretion parameters were measured inDictyostelium discoideum amoebae constructed from strain AX3 by transformation with a multicopy plasmid carrying either a normalras gene (ras-Gly12), a mutatedras gene (ras-Thr12) or by the vector carrying the geneticin resistance gene only (pDNEO2). It was found that the pinocytosis rate and extent as well as the lysosomal enzyme secretion were slightly different in the three strains. These changes, however, were related to minor modifications of the cellular volumes. The overall concentration of inositol hexakisphosphate was similar in the three strains.This work was supported in part by a grant from the Ligue Nationale Française contre le Cancer (n^o 880258) to MS, and by a grant from the Fonds National Suisse de la Recherche Scientifique (No. 3.623-087) to CR.     

Esterase activity in the hepatopancreas ofMacrobrachium lamarrei (Crustacea: Decapoda)

Summary The activity of the hepatopancreatic esterase of the fresh water prawnMacrobrachium lamarrei was optimal at pH 7.4 and temperature 40°C. The activity increased with the increase in incubation period and enzyme concentration. The Michaelis constant (K_m) of the enzyme was 2.1×10^–3M.The investigations are a part of the thesis presented to University of Lucknow, India.Acknowledgments. The authors are grateful to the University Grants Commission, India for the award of a Junior Research Fellowship.     

Effect of lectins from leguminous seeds on rat duodenal enterokinase activity

Summary Enterokinase activity from rat duodenal brush borders was assayed in vitro in the presence of purified lectins from 3 leguminous seeds. Noncompetitive inhibition of the enzyme was observed in each case.Phaseolus hemagglutinin was the most potent inhibitor among the 3 lectins tested.     

Propriétés antiacétylcholinestérasiques du diiodométhylate debis-(diméthylamino-3-phénoxy)-1-3 propane,de ses dérivés phénoliques et des esters carbamiques correspondants

Summary Thein vitro anti-acetylcholinesterase properties ofbis-(dimethylamino-3-phenoxy)-1-3 propane dimethiodide (2842 CT) of two phenolics derivatives (3443 CT and 3116 CT) and of the two corresponding carbamic esters (3152 CT et 3113 CT) have been compared using human red blood corpuscles as enzyme source; under specified conditions, the Cl-50 are respectively 8 × 10^–7 M for 2842 CT, 3.5 × 10^–9 for the two phenolic compounds, and 1.5 × 10^–9 for the carbamic esters. The potencies of these phenols are very close to those of the carbamates, being a bit higher or lower depending on the concentration of the inhibitor and on the time of the readingThe two phenolic compounds, like 2842 CT, react readily with the enzyme contrarily to the carbamic esters which combine slowly. On the other hand the inhibition by the phenolic derivatives is as stable against washing as that by the carbamates. The carbamates, but not the phenols, show the slow displacement phenomenon.Some of these characteristics are compatible with the hypothesis that carbamic compounds could act through liberated phenolic functions but others indicate that carbamic groups have a role of their own.     

Aktivierung einer alkalischen Phosphatase

Summary -Glycerophosphatase prepared from the intestinal mucosa of the calf was purified by fractionated precipitation with alcohol. A further concentration of the enzyme activity was attained by electrophoresis.The activity of the purified enzyme solution was reduced to of its original value when dialysed for 48 hours atp _H 4.5. Atp _H 6 and atp _H 10.5 only a less pronounced decrease of the activity occurred.By addition of heat-inactivated-glycerophosphatase to the enzyme solution which was partly inactivated by dialysis atp _H 4.5 the activity of the latter was increased by about 100%.     

Über das Vorkommen und die Natur der Cholinesterase der Schlangengifte

Summary An enzyme that hydrolyzes acetylcholine is present in the dried venoms of 19 species of theColubridae (9 genera), but not in the venoms of more than 20 species of theViperidae (7 genera). The enzyme behaves like the e-Cholinesterase of the mammalian tissues.     

Mitochondrial DNA variation in geographic populations of Colorado potato beetle,Leptinotarsa decemlineata (Coleoptera; Chrysomelidae)

Summary This study demonstrates variability in restriction enzyme cleavage sites of mitochondrial DNA (mtDNA) among four popalations of Colorado potato beetle (CPB). A suite of three enzymes (EcoRI,HpaI,PstI) was sufficient to discriminate among the populations tested. Individuals heteroplasmic for restriction enzyme patterns were found in some populations. Variability in CPB mtDNA should prove useful in efforts to trace the origin and dispersal of the species in North America.     

Inhibitory effect of flavonoids on DNA-dependent DNA and RNA polymerases

Summary Flavonoids, (–)-epigallocatechin (1), myricetin (2) and quercetin (3), were investigated for inhibitory effects onE. coli DNA polymerase I and T7 bacteriophage RNA polymerase. In both DNA and RNA synthesis,1 and3 inhibited enzyme reactions by non-competitive and mixed type inhibition respecitively, with regard to template DNAs. Myricetin (2) inhibited DNA and RNA polymerase reactions by mixed type and competitive type inhibition, respectively, with template DNAs. It was suggested that2 interacts with covalently closed/circular DNA.     

Monoclonal antibodies to L-asparaginase

Summary Five hybridomas secreting monoclonal antibody toE. coli L-asparaginase were isolated. These monoclonal antibodies were classified into 3 different subclasses; Ig G₁ (1 clone), Ig G₂ (2 clones) and Ig G₃ (2 clones). One of them possessed anti-L-asparaginase neutralizing activity. Four antibodies examined demonstrated a linear Langmuir binding plot and binding affinities, with equilibrium dissociation constant (K_d) ranging between 2.5×10^–9M and 6.3×10^–10 M. The monoclonal antibodies should be useful probes for investigation of the enzyme activity.