Transformation of some hydroxy amino acids to other amino acids.

It has been observed that beta-hydroxy-alpha-amino acids are transformed into other amino acids, when heated in dilute solutions with phosphorous acid, phosphoric acid or their ammonium salts. It has been shown that as in the case of previously reported glycine-aldehyde reactions, glycine also reacts with acetone to give beta-hydroxyvaline under prebiologically feasible conditions. It is suggested, therefore, that the formation of beta-hydroxy-alpha-amino acids and their transformation to other amino acids may have been a pathway for the synthesis of amino acids under primitive earth conditions.     

Modeling amino acid replacement.

The estimation of amino acid replacement frequencies during molecular evolution is crucial for many applications in sequence analysis. Score matrices for database search programs or phylogenetic analysis rely on such models of protein evolution. Pioneering work was done by Dayhoff et al. (1978) who formulated a Markov model of evolution and derived the famous PAM score matrices. Her estimation procedure for amino acid exchange frequencies is restricted to pairs of proteins that have a constant and small degree of divergence. Here we present an improved estimator, called the resolvent method, that is not subject to these limitations. This extension of Dayhoff's approach enables us to estimate an amino acid substitution model from alignments of varying degree of divergence. Extensive simulations show the capability of the new estimator to recover accurately the exchange frequencies among amino acids. Based on the SYSTERS database of aligned protein families (Krause and Vingron, 1998) we recompute a series of score matrices.     

氨基酸-山梨醇输液的研制

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Accumulation of amino acids by lysosomes incubated with amino acid methyl esters.

Rat liver lysosomal preparations incubated with 10(-5) M L-[4,5-3H]leucine methyl ester hydrolyzed the methyl ester and accumulated radioactivity within a particulate compartment. The acculated radioactivity was identified as free leucine by thin layer chromatography. Free leucine was not itself taken up by the lysosomal preparations. The capacity to accumulate leucine was identified as a specific property of lysosomes and was thought to result from the trapping of the free amino acid within the lysosome following the hydrolysis of the methyl ester. Lysosomes also accumulated phenylalanine, serine, and alanine when incubated with the corresponding methyl esters. Leucine accumulation was inhibited by submillimolar concentrations of chloroquine, by the protease inhibitor L-1-tosylamido-2-phenylethyl chloromethyl ketone, and by lowering the pH below 7.0. Efflux of leucine from the lysosomes was highly temperature dependent (activation energy 33 kcal/mol). No evidence was found to suggest that leucine efflux was a carrier-mediated process. The results provide a new methodology for the study of amino acid movements across lysosomal membranes.     

Ion-exchange chromatography of sulfur amino acids on a single-column amino acid analyzer.

Examination of the extent of production of the ninhydrin-colored derivative, Ruhemann's purple, under automated conditions of a single-column amino acid analyzer by several classes of sulfur-containing amino acids revealed a wide variation in the color factors relative to leucine. These ranged from 0.02 for the methyl ester of cysteine to 2.19 for D-homocystine. Color yields obtained by the manual ninhydrin reaction are generally lower than the corresponding values obtained on the amino acid analyzer. The elution positions ranged from 5.12 min for cysteic acid to 84.9 min for l-cystine dimethyl ester. The observed behavior of these compounds in the ninhydrin reaction is rationalized in terms of structural and electronic factors which they exhibit in reacting with ninhydrin to form the visible dye. Such an analysis should make it possible to predict ninhydrin color factors, and possibly also elution times, of structurally related compounds.     

Polycondensation of thermal precursors of amino acids and characterization of constituent amino acids.

As a model reaction of polyamino acid formation from non-amino acid precursors, diammonium citraconate (I), ammonium citraconamate(II) and ammonium itaconamate(III) were converted to polyimide type polymers by thermal polycondensation by heating at 130–210°C. The imide type polymer was partially hydrolyzed to the corresponding peptide type polyamino acid. The polymer was composed of α-methylaspartic acid (IV), threo- and erythro-ß-methylaspartic acid (V) and α-(aminomethyl) succinic acid (VI). On the other hand, IV was thermally polycondensed to the corresponding polymer. It was found that the amino acid composition of the polymer was similar to that of the polymer prepared from I, II and III. The formation and isomerization of amino acids during the thermal polycondensation are described.     

Fullerene-based amino acids and peptides.

Recent advances in the chemistry of fullerene have allowed the synthesis of many classes of novel fullerene derivatives. Among these classes, fullerene-based amino acids and peptides are particularly interesting, both for structural studies and biological applications. In this review, we will discuss our own achievements in this rapidly growing field. In particular, the application of fulleroproline (Fpr) amino acids and peptides to medicinal chemistry and material science will be highlighted.     

Permeability of amino acids into liposomes.

1. A simple and rapid assay for the measurement of permeability of amino acids into liposome membrane was carried out by using the liposomes trapping D-amino acid oxidase (D-amino acid: O2 oxidoreductase (deaminating), EC 1.4.3.3) inside the membrane. 2. Permeability of amino acids into liposomes depended on the lipid composition of the membrane. Permeability of amino acids into phosphatidylcholine-cholesterol liposomes depended critically on temperature. 3. Permeability also depended on the structure of amino acids. The order of permeability was norvaline greater than isoleucine greater than leucine greater than phenylalanine greater than tryptophan greater than methionine greater than tyrosine, valine greater than threonine greater than serine greater than alanine greater than glycine.     

E.s.r. of spin-trapped radicals in gamma-irradiated polycrystalline amino acids, N-acetyl amino acids and dipeptides

The radicals produced in several polycrystalline amino acids, N-acetyl amino acids and dipeptides by gamma-radiolysis at room temperature were investigated by spin-trapping. After irradiation in the solid state, the samples were dissolved in aqueous solutions f t-nitrosobutane and the trapped radicals identified by e.s.r. For alpha-amino acids, deamination radicals were found, and in some cases H-abstraction radicals were also observed. No decarboxylation radicals could be detected. For N-acetyl amino acids, except for N-acetylglycine, the major radical was the decarboxylation radical. For N-acetyglycine the H-abstraction radical from the glycine residue was observed. For dipeptides of the x-glycine, the radical formed by removal of H from the alpha-carbon of the carboxyl-terminal residue was always spin-trapped. Some primary deamination radicals and minor amounts of decarboxylation radicals could also be observed. For dipeptides of the type x-alanine, glycine-x and alanine-x, the decarboxylation radical was always the major spin-trapped radical. Some primary and secondary deamination radicals were also detected.     

Derepression of amino acid transport by amino acid starvation in rat hepatoma cells.

Amino acid starvation causes an adaptive increase in the initial rate of transport of selected neutral amino acids in an established line of rat hepatoma cells in tissue culture. After a lag of 30 min, the initial rate of transport of alpha-aminoisobutyric acid (AIB) increases to a maximum after 4 to 6 h starvation of 2 to 3 times that seen in control cells. The increased rate of transport is accompanied by an increase in the Vmax and a modest decrease in the Km for this transport system, and is reversed by readdition of amino acids. The enhancement is specific for amino acids transported by the A or alanine-preferring system (AIB, glycine, proline); uptake of amino acids transported by the L or leucine-preferring system (threonine, phenylalanine, tyrosine, leucine) or the Ly+ system for dibasci amino acids (lysine) is decreased under these conditions. Amino acids which compete with AIB for transport also prevent the starvation-induced increase in AIB transport; amino acids which do not compete fail to prevent the enhancement. Paradoxically threonine, phenylalanine, tryptophan, and tyrosine, which do not compete with AIB for transport, block the enhancement of transport upon amino acid starvation. The starvation-induced enhancement of amino acid transport does not appear to be the result of a release from transinhibition. After 30 min of amino acid starvation, AIB transport is either unchanged or slightly decreased even though amino acid pools are already depleted. Furthermore, loading cells with high concentrations of a single amino acid following a period of amino acid starvation fails to prevent the enhancement of AIB transport, whereas incubation of the cells with the single amino acid for the entire duration of amino acid starvation prevents the enhancement; intracellular amino acid pools are similar under both conditions. The enhancement of amino acid transport requires concomitant RNA and protein synthesis, consistent with the view that the adaptive increase reflects an increased amount of a rate-limiting protein involved in the transport process. Dexamethasone, which dramatically inhibits AIB transport in cells incubated in amino acid-containing medium, both blocks the starvation-induced increase in AIB transport, and causes a time-dependent decrease in transport velocity in cells whose transport has previously been enhanced by starvation.     

T.l.c. enantiomeric separation of amino acids

Resolution of enantiomers is very important particularly in the fields of asymmetric synthesis, mechanistic studies, geochronology, studies of structure-function relationship of proteins, pharmacology, and medicine. Various chromatographic methods have replaced the classical fractional crystallization, seeding and enzymatic procedures. Of these, t.l.c. provides a direct, simple, and inexpensive method for resolution of enantiomers of amino acids and their derivatives. Ligand exchange, ion exchange, and molecular inclusion complexation have been the basis of t.l.c. resolution of enantiomers of amino acids and their derivatives. The innovation of new plate types, and methods of development and detection have renewed interest in the direct resolution of enantiomers of amino acids, their derivatives and a variety of other compounds by t.l.c. The present report provides an overview of some of the more recent approaches to the direct t.l.c. resolution of amino acids and their derivatives together with special advantages and scope of t.l.c.