Genetisch bedingte Tumoren und der Gehalt an freien Aminosäuren beiNicotiana

Summary The amphidiploid hybrid ofNicotiana glauca ×N. langsdorffi, producing tumors spontaneously, contains essentially more of the following free amino acids than the inital species: cystine, histamine, oxyproline, alanine, kynurenine, tryptophane, and a non-identified ninhydrine positive substance. It is supposed that the abnormally high content of free amino acids is connected with the spontaneous formation of tumors.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Conformational constraints on side chains in protein residues increase their information content

Like all other complex biological systems, proteins exhibit properties not seen in free amino acids (i.e., emergent properties). The present investigation arose from the deduction that proteins should offer a good model to approach the reverse phenomenon, namely top-down constraints experienced by protein residues compared to free amino acids. The crystalline structure of profilin Ib, a contractile protein of Acanthamoeba castellanii, was chosen as the object of study and submitted to 2-ns molecular dynamics simulation. The results revealed strong conformational constraints on the side chain of residues compared to the respective free amino acids. A Shannon entropy (SE) analysis of the conformational behavior of the side chains showed in most cases a strong decrease in the SE of the 1 and 2 dihedral angles compared to free amino acids. This is equivalent to stating that conformational constraints on the side chain of residues increase their information content and hence recognition specificity compared to free amino acids. In other words, the vastly increased information content of a protein relative to its free monomers is embedded not only in the tertiary structure of the backbone, but also in the conformational behavior of the side chains. The postulated implication is that both backbone and side chains, by virtue of being conformationally constrained, contribute to the recognition specificity of the protein toward other macromolecules and ligands.Received 13 July 2003; received after revision 18 August 2003; accepted 4 September 2003     

Characterization of free amino acids in the hemolymph ofAchaea janata L. larvae (Lep.Noctuidae)

Summary 19 free amino acids were separated and quantified from the hemolymph of 5th instar larvae of the mothAchaea janata. Proline, histidine, threonine/glutamine/asparagine, lysine, valine and serine were the predominant amino acids in the hemolymph. Changes in amino acid concentration are discussed in relation to metabolic and other physiological activities.Acknowledgment. Y.P. Ramdev wishes to thank the Director, Indian Agricultural Research Institute, New Delhi, India for providing Senior Research Fellowship.     

Free amino acids in some tissues of marine crustacea

Summary Free amino acids contained in extracts of crustaceans were separated and determined quantitatively by ion exchange chromatography. In the hepatoprancreas of the brachyuran crustaceanCarcinus maenas, glycine, arginine, proline and alanine constituted 78.7% of the total amino acid pool. InEriphia spinifrons andMaja verrucosa, glycine, alanine, lysine and leucine comprised 54.9–66.9% of the total content. In the muscle tissue of the macruran crustaceaPalaemon, glycine, arginine, proline and alanine were the most common free amino acids and comprised 70–95% of the total content. The high concentrations of some amino acids in tissues of crustaceans may play a role in intracellular osmotic regulation and also in the maintenance of ionic hydrogen concentration.Acknowledgments. The author wishes to thank Mr R. Mari for his skillful help in the amino acid analyses and Miss G. Princivalli for typing the mansucript.     

Free amino acids in the adult citrus brown mite,Eutetranychus orientalis (Klein)

Summary Amino acids contained in extracts of adultEutetranychus orientalis were separated and determined quantitatively by 2-dimensional paper chromatography. 14 amino acids were identified. Asparagine, ornithine, histidine, lysine, aspartic acid, serine and glycine were the major components of the free amino acid pool, comprising 83.94% of the total content.     

Lysine estimation with the modified Udy-dye binding method in hexaploid wheat

Summary Lysine content in bread wheat (Triticum aestivum L. m. Thell) was determined by a modified Udy-dye binding method and was compared with that obtained from the amino acid analyzer. The values obtained from the 2 methods were correlated and the co-efficient was found to be +0.91 at 0.450 mg/ml dye concentration. The modified method is quick, less expensive and quite helpful for screening lysine at earlier generation in wheat breeding for improved grain quality content.The author is grateful to Dr V.A. Johnson, University of Nebraska, Lincoln, USA, for supplying wheat cultivars with known protein and lysine values.     

Epigenetic mechanisms in mammals

DNA and histone methylation are linked and subjected to mitotic inheritance in mammals. Yet how methylation is propagated and maintained between successive cell divisions is not fully understood. A series of enzyme families that can add methylation marks to cytosine nucleobases, and lysine and arginine amino acid residues has been discovered. Apart from methyltransferases, there are also histone modification enzymes and accessory proteins, which can facilitate and/or target epigenetic marks. Several lysine and arginine demethylases have been discovered recently, and the presence of an active DNA demethylase is speculated in mammalian cells. A mammalian methyl DNA binding protein MBD2 and de novo DNA methyltransferase DNMT3A and DNMT3B are shown experimentally to possess DNA demethylase activity. Thus, complex mammalian epigenetic mechanisms appear to be dynamic yet reversible along with a well-choreographed set of events that take place during mammalian development.     

Free amino acids of the haemolymph of the cotton leaf-worm,Spodoptera littoralis Boisduval full-grown larvae,infected with nuclear-polyhedrosis virus

Summary Free amino acids in the haemolymph ofSpodoptera littoralis full-grown larvae infected with a nuclearpolyhedrosis virus were compared with those in the haemolymph of normal insects. Amino acids were separated by 2-dimensional paper chromatography and quantified colorimetrically. Most of the amino acids in the haemolymph of diseased full-grown larvae decreased markedly in concentration but proline, lysine, aspartic acid and histidine occurred in greater concentration in the haemolymph of diseased full-grown larvae than in the haemolymph of healthy insects.     

Synthesis, structure and function of poly-α-amino acids – the simplest of protein models

During the 1950s, linear and multichain poly-α-amino acids were synthesized by polymerization of the corresponding N-carboxyamino acid anhydrides in solution in the presence of suitable catalysts. The resulting homo- and heteropolymers have since been widely employed as simple protein models. Under appropriate conditions, poly-α-amino acids, in the solid state and in solution, were found to acquire conformations of an α-helix and of β-parallel and antiparallel pleated sheets, or to exist as random coils. Their use in experimental and theoretical investigations of helix-coil transitions helped to shed new light on the mechanisms involved in protein denaturation. Conformational fluctuations of peptides in solution were analysed theoretically and studied experimentally by nonradiative energy-transfer techniques. Poly-α-amino acids played an important role in the deciphering of the genetic code. In addition, analysis of the antigenicity of poly-α-amino acids led to the elucidation of the factors determining the antigenicity of proteins and peptides. The synthetic procedures developed made possible the preparation of immobilized enzymes which were shown to be of considerable use as heterogeneous biocatalysts in the chemical and pharmaceutical industry. Interest in the biological and physicochemical characteristics of poly-α-amino acids was recently renewed because of the reported novel findings that some copolymers of amino acids are effective as drugs in multiple sclerosis, and that glutamine repeats and reiteration of other amino acids occur in inherited neurodegenerative diseases.     

Quelques aspects de la croissance rhizomorphique chez leSphaerostilbe repens

Summary The rhizomorphs ofSphaerostilbe repens attached to synnemata and mycelia or severed from them, grew providing that their apices were kept inside the culture medium. Exposed to air, apical regions became dark brown, stopped growing and differentiated into synnemata. However central canals in the middle of the rhizomorphs had to be continuously maintained in contact with the atmosphere; nitrogen immediately stopped rhizomorphic growth. This indicates that oxygen for growing apices is supplied through central canals and that the partial pressure of oxygen has to be higher at the origin of the rhizomorphs than that which is in contact with the outside surfaces of their apices.     

On the significance of amino acids in the larval development of khapra-beetle,Trogoderma granarium everts. (Coleoptera: Dermestidae)

Résumé On décrit un milieu de composition chimique définie, se prêtant à l'analyse qualitative des acides aminés nécessaires au développement des larves deTrogoderma. A cet effet, dix acides aminés sont indispensables: l'arginine, l'histidine, l'isoleucine, la leucine, la lysine, la méthionine, la phénylalanine, la thréonine, la tryptophane et la valine. Le lard et l'acide nucléique sont également nécessaires à l'alimentation de ces larves.     

Conotoxins and the posttranslational modification of secreted gene products

The venoms of predatory cone snails (genus Conus) have yielded a complex library of about 50–100,000 bioactive peptides, each believed to have a specific physiological target (although peptides from different species may overlap in their target specificity). Conus has evolved the equivalent of a drug development strategy that combines the accelerated evolution of toxin sequences with an unprecedented degree of posttranslational modification. Some Conus venom peptide families are the most highly posttranslationally modified classes of gene products known. We review the variety and complexity of posttranslational modifications documented in Conus peptides so far, and explore the potential of Conus venom peptides as a model system for a more general understanding of which secreted gene products may have modified amino acids. Although the database of modified conotoxins is growing rapidly, there are far more questions raised than answers provided about possible mechanisms and functions of posttranslational modifications in Conus. Received 24 June 2005; received after revision 13 August 2005; accepted 19 September 2005     

Changes in free amino acid level due to physical and chemical agents in ageingDrosophila

Summary The effects of gamma rays and caffeine on total free amino acids were assessed in unirradiated and irradiated ageingDrosophila. As age advances, there is a rise in the quantity of free amino acids in untreated males and females. Gamma rays and caffeine strongly reduce the level of free amino acids in both the sexes.Acknowledgment. The author wishes to thank the Dean, School of life Sciences, Jawaharlal Nehru University, New Delhi-110067, for providing necessary fascilities during the course of this work.     

Inhibition of arginase in sheep brain homogenates by some L-amino acids

Summary The effect of 16 L-amino acids on the activity levels of arginase in sheep brain homogenates was studied. The amino acids leucine, valine, lysine, and ornithine inhibited arginase activity significantly. The other amino acids tested did not show a significant influence on arginase activity. The inhibition was related to the carbon chain length of the amino acids.     

"Add-on" domains of<Emphasis Type="Italic"> Drosophila</Emphasis> β1,4-<Emphasis Type="Italic">N</Emphasis>-acetylgalactosaminyltransferase B in the stem region and its pilot protein

The glycolipid specific Drosophila melanogaster β1,4-N-acetylgalactosaminyltransferase B (β4GalNAcTB) depends on a zinc finger DHHC protein family member named GalNAcTB pilot (GABPI) for activity and translocation to the Golgi. The six-membrane spanning protein actually lacks the cysteine in the cytoplasmic DHHC motif, displaying DHHS instead. Here we show that the whole conserved region around the DHHS sequence, which is essential for palmitoylation in DHHC proteins, is not required for GABPI to interact with β4GalNAcTB. In contrast, the two luminal loops between transmembrane domain 3–4 and 5–6 contain conserved amino acids, which are crucial for activity. Besides the dependence on GABPI, β4GalNAcTB requires its exceptional short stem region for activity. A few hydrophobic amino acids positioned close to the transmembrane domain are essential for the interaction with GABPI. Along with its catalytic domain, β4GalNAcTB, thus, requires an area in its own stem region and two small luminal loops of GABPI as "add-on" domains. Moreover, some inactive GABPI mutants could be rescued by fusion with β4GalNAcTB, indicating their importance in direct GABPI-β4GalNAcTB interaction.     

Genetische und biochemische Untersuchungen über die Bedeutung der Amino- und Nucleinsäuren im Ursachengefüge von Neoplasmen (Tumoren und Gallen). Ein Dauermodifikationsbzw. Prädeterminationsphänomen

Summary If certain hybrids ofPlatypoecilus maculatus andXiphophorus helleri (viviparous platyfish and swordtails) are cultivated in diluted sea-water (0.25 or 0.5% salt concentration), the amount of free amino acids increases and the growth of macromelanophores, which normally only form certain black spots at the dorsal fin, is accelerated. Therefore melanomas arise. After changing salt water for fresh water, growth of macromelanophores remains accelerated. The same effect is seen in progenies which have been in an oocytic or embryonic state, when their mother was cultivated for some weeks in diluted sea-water (Figures 1 and 2). A similar effect has been observed in plant galls produced byPhylloxera (Viteus vitifolii) in the genusVitis. When the parasite, which injects amino acids into the tissue of the host in order to produce galls, is removed, gall formation goes on slowly for some days. It is shown that a few hours after injection of amino acids—before formation of neoplasms begins—the concentration of ribonucleic acids is increased. RNS-(and therefore protein-)biosynthesis in neoplasms may be dependent on concentration of free amino acids. A simple concept for the formation of neoplasms in hybrids of platyfish and galls ofPhylloxera is discussed.

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Den Herren Prof. Dr.G. de Lattin (Zoologisches Institut der Universität des Saarlandes) und Prof. Dr.B. Husfeld (Forschungsinstitut für Rebenzüchtung Geilweilerhof/Pfalz) sind wir für die Förderung dieser Arbeit zu grösstem Dank verpflichtet. Herrn Prof. Dr.C. Kosswig (Zoologisches Institut der Universität Hamburg) verdanken wir viele Anregungen und die freundliche Überlassung von ingezüchteten Stämmen für unsere Fischkreuzungen. — Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Benützung der oszillographischen Polarographie in der Mikrobiologie

Summary Oscillographic polarography with alternating current was used for following the course of utilisation and metabolism of some amino acids, purine and pyrimidine bases, similarly as their nucleosides and nucleotides in the washed suspensions of resting cells and in growing cultures ofEscherichia coli. The Polaroscope P-524 was used with mercury dropping electrode. The method was described for the oscillopolarographic investigation of activity of some bacterial enzymes, as for example: asparaginase, some nucleodesaminases and enzymes involved in the cleavage of nucleosides.     

Incorporation d'acides aminés radioactifs dans les membranes des globules lipidiques du lait humain

Summary Human milk fat globule membranes (MFGM) can incorporate radioactive¹⁴C amino acids in a hot trichloracetic acid-insoluble material. Aspecific adsorption and bacterial contamination are unlikely. The products of protein synthesis were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate or by action of proteolytic enzymes. Various inhibitors of protein synthesis were assayed. Fragments of rough endoplasmic reticulum or mitochondria could be involved in this incorporation.     

Alternative exon usage selectively determines both tissue distribution and subcellular localization of the acyl-CoA thioesterase 7 gene products

Acyl-CoA thioesterases (ACOTs) catalyze the hydrolysis of acyl-CoAs to free fatty acids and coenzyme A. Recent studies have demonstrated that one gene named Acot7, reported to be mainly expressed in brain and testis, is transcribed in several different isoforms by alternative usage of first exons. Strongly decreased levels of ACOT7 activity and protein in both mitochondria and cytosol was reported in patients diagnosed with fatty acid oxidation defects, linking ACOT7 function to regulation of fatty acid oxidation in other tissues. In this study, we have identified five possible first exons in mouse Acot7 (Acot7a–e) and show that all five first exons are transcribed in a tissue-specific manner. Taken together, these data show that the Acot7 gene is expressed as multiple isoforms in a tissue-specific manner, and that expression in tissues other than brain and testis is likely to play important roles in fatty acid metabolism. Received 5 February 2007: received after revision 3 April 2007; accepted 19 April 2007     

A human homolog of the yeast gene encoding tRNA 2′-phosphotransferase: cloning,characterization and complementation analysis

The Saccharomyces cerevisiae TPT1 gene plays a role in removing the 2-phosphate from ligated tRNA during the maturation of pre-tRNA. Here we reported the cloning and characterization of the human TRPT1 gene as a homolog of yeast TPT1. The TRPT1 gene is located at human chromosome 11q13 and encodes a polypeptide of 253 amino acids. BLAST searches with its amino acid sequence revealed the ubiquitous occurrence of TRPT1 homologs and their functional relationships with the presence of the DUF60/KptA domain. Northern analysis demonstrated that the gene is primarily expressed in heart and skeletal muscle, with lower or undetectable levels in other tissues studied. A plasmid-shuffling experiment showed that the human TRPT1 gene could complement the tpt1 mutation in S. cerevisiaeReceived 19 March 2003; received after revision 25 April 2003; accepted 22 May 2003