不同晶型、旋光型氨基酸原料药的红外图谱分析

分析比较了 33种不同来源的氨基酸产品红外图谱的差异 ,其中丝氨酸、门冬氨酸、醋酸赖氨酸、谷氨酸 (白色结晶性粉末 )、苏氨酸、缬氨酸、丙氨酸、亮氨酸、脯氨酸、盐酸组氨酸、盐酸精氨酸、酪氨酸、胱氨酸等 13种与标准图谱完全一致 ;甲硫氨酸、盐酸赖氨酸、甘氨酸、谷氨酸 (白色结晶 )等 4种与标准图谱不一致 ,其原因是 :甘氨酸和谷氨酸由晶型不同造成 ,甲硫氨酸因旋光性不同而造成 ,盐酸赖氨酸与相应的生化试剂图谱一致。     

Relationship between the in vivo and in vitro activity of some naturally occurring glutamate analogues on the somatic neuromuscular junction of Lucilia sericata

ABSTRACT. A preparation of Lucilia sericata flight motor system was arranged so that ganglionic and neuromuscular function could be monitored while experimental compounds were injected into the intact insect. Injections of l -glutamate, the putative excitatory transmitter at the insect neuromuscular junction, caused a reversible paralysis of the flight muscles. A number of structural analogues of l -glutamic acid, found in various seed plants, were injected and the results compared. The salts of several of these compounds were as active or more active in causing the paralysis than glutamate itself. Two of the most toxic compounds, salts of 4-methylene glutamic acid and quisqualic acid were further tested in vitro by iontophoretically applying them directly to exposed insect neuromuscular junctions. Both compounds showed glutamate-agonistic activity when applied directly to the neuromuscular junction but were less active than glutamate. This difference between in vivo and in vitro effects is caused by removal mechanisms which protect the muscle membranes from the effects of glutamate. These mechanisms do not so readily remove or inactivate 4-methylene glutamate or quisqualate. Consequently, for a given dose, the concentration of the analogues at the neuromuscular junction remains longer above the critical level which causes paralysis.     

Specificity of Amino Acid Transport in Trypanosoma equiperdum*

Uptake of [¹⁴C] alanine, arginine, glutamic acid and phenylalanine by Trypanosoma equiperdum occurred by both a mediated mechanism and diffusion. Twenty amino acids were studied as inhibitors of absorption of the above amino acids. Results suggested that at least 4 distinct transport loci are involved in amino acid transport. These 4 loci have overlapping affinities for amino acids and seem to be involved, respectively, in the absorption of (a) arginine and phenylalanine; (b) arginine; (c) alanine, phenylalanine, and glutamic acid; (d) glutamic acid. The data also showed that multiple sites for substrate binding occur on each of 2 transport systems.     

Specificity of aspartate aminotransferases from leguminous plants for 4-substituted glutamic acids

Aspartate aminotransferase (glutamate-oxalacetate transaminase) was partially purified from extracts of germinating seeds of peanut (Arachis hypogaea), honey locust (Gleditsia triacanthos), soybean (Glycine max), and Sophora japonica. The ability of these enzyme preparations, as well as aspartate aminotransferase purified from pig heart cytosol, to use 4-substituted glutamic acids as amino group donors and their corresponding 2-oxo acids as amino group acceptors in the aminotransferase reaction was measured. All 4-substituted glutamic acid analogs tested were poorer substrates than was glutamate or 2-oxoglutarate. 2-Oxo-4-methyleneglutarate was least effective (lowest relative V_m/K_m) as a substrate for the enzyme from peanuts and honey locust, which are the two species studied that accumulate 4-methyleneglutamic acid and 4-methyleneglutamine. Of the different aminotransferases tested, the enzyme from honey locust was the least active with 2-oxo-4-hydroxy-4-methylglutarate, the corresponding amino acid of which also accumulates in that species. These results suggest that transamination of 2-oxo-4-substituted glutaric acids is not involved in the biosynthesis of the corresponding 4-substituted glutamic acids in these species. Rather, accumulation of certain 4-substituted glutamic acids in these instances may be, in part, the result of the inefficacy of their transamination by aspartate aminotransferase.     

Effect of nicotine on the level of extracellular amino acids in the hippocampus of rat

Using microdialysis, we compared intracerebral and subcutaneous administration of nicotine for the effect on the levels of extracellular amino acids in the hippocampus of anesthetized rats. Administration by microdialysis of 10 mM nicotine, resulting in a nicotine concentration of 0.134 μmol/g in the hippocampus, increased the extracellular levels of aspartic acid, glutamic acid, and serine by 26–60%. At 50 mM nicotine the increases in the levels of aspartic acid, glutamic acid, serine, glycine, and glutamine were between 76% and 141%. Subcutaneous administration of nicotine at a dose of 6 μmol/kg caused a 57% increase in the extracellular level of glutamic acid. After a dose of 12 μmol/kg that resulted in a nicotine level of 0.015 μmol/kg in the hippocampus, the extracellular level of glutamic acid was increased by 100%, and that of aspartic acid by 24%. Thus, higher cerebral nicotine levels were needed with intracerebral than with subcutaneous administration to obtain similar amino acid changes. Prior administration of mecamylamine or L-kynurenine prevented the subcutaneous nicotine-induced elevation of the extracellular levels of aspartic acid and glutamic acid. Our results indicate that receptor interactions modulate nicotine effects and that both nicotinic cholinergic and NMDA/glycine glutamatergic receptors participate in the action of nicotine in increasing extracellular amino acid levels.     

Variations in the content of free and bound amino acids during dormancy and the first cell cycle in Helianthus tuberosus tuber explants

Abstract

Qualitative and quantitative analysis of free and bound amino acids and amides during dormancy and the most important phases of the first cell cycle was carried out in tubers of Helianthus tuberosus.

In the dormant tuber arginine was confirmed to be the most abundant amino acid. A high amount of asparagine was also present; on the contrary glutamine was found in very low concentrations. During the progression of dormancy, all the free amino acids and amides declined while aspartic and glutamic acid increased.

During the G₁ phase of the first cell cycle induced by 2,4-D, all the free amino acids and amides decreased with the exception of glutamic acid.

At 18, 20, 24 h of activation with 2,4-D, corresponding to the S phase and the beginning of mitosis, bound amino acids were also determined. In these phases of the cell cycle they increased reaching a maximum at 20 h; on the other hand the free amino acid and amide content, especially aspartic acid, asparagine and arginine, decreased with the exception of glutamic acid, alanine and phenylalanine.     

The Molecular Biology of Euglena gracilis IX. Amino Acid Pool Composition

The amino acid composition of the acid soluble fraction of Euglena gracilis was determined from cells grown in 4 different culture media. Glutamic acid is the major free amino acid. Hydrolysis of this fraction increases the amount of free amino groups, the major amino acids found are then glutamic acid, aspartic acid, glycine and arginine. The pattern of amino acid distribution is similar in all 4 culture media. L-arginyl-L-glutamine was isolated and identified in extracts from all 4 culture conditions. It was shown to be a metabolic intermediate by radioactivity chase experiments.     

Mass Fragmentographic Method for the Determination of Trace Amounts of Putative Amino Acid Neurotransmitters and Related Compounds from Brain Perfusates Collected In Vivo

Abstract: A mass fragmentographic method for the determination of trace amounts of amino acid neurotransmitter candidates from brain perfusates is described. The analytical procedure includes the measurements of glycine, β-alanine, γ-aminobutyric acid, proline, aspartic acid, and glutamic acid; αalanine, leucine, and sarcosine, undergoing gas chromatographic coelution, are detected simultaneously. Amino acids extracted from dried perfusate residues are converted to the corresponding N -pentafluoropropionyl hexafluoroisopropyl esters by a single-step procedure. Gas chromatographic separation of the amino acid derivatives is achieved on a packed glass column filled with trifluoropropylsilicone as stationary phase. The limit of detection for the different derivatives (signal-to-noise, 3:1) ranges from 50 femtomol to 1 picomol. Deuterium-labeled amino acid analogues are used as internal standards for quantitative measurements. The mass spectral characteristics of the derivatives are compared and discussed. The technique has been applied to the assay of amino acids released in vivo within the pigeon optic tectum, demonstrating the capabilities of the present analytical approach.     

Increasing dispensable amino acids in diets of kittens fed essential amino acids at or below their requirement increases the requirement for arginine

Summary Kittens fed diets containing 0.75 × the NRC (1986) essential amino acid requirement (EAArq) and 210 to 560g crude protein(CP)/kg diet exhibited, with increasing CP: 1) decreasing weight gain, 2) decreasing plasma arginine concentrations, 3) increasing urinary orotic acid excretion, 4) increasing plasma glutamic acid concentrations, and 5) plasma isoleucine concentrations at levels that suggest a marginal isoleucine deficiency. Kittens fed a control diet (CD) containing 1.5 × EAArq and 350 g CP/kg diet had maximal weight gains and no orotic aciduria. It was concluded that the decreased weight gain and adverse metabolic effects were caused by arginine deficiency and possibly glutamic acid toxicity induced by high dietary dispensable amino acids. Kittens fed the diets containing 1.0 × EAArq and 350 and 560 g CP/kg diet had depressed plasma arginine and elevated glutamic acid concentrations and orotic aciduria. These results indicate that 10 g arg/kg diet is not adequate at CP concentrations above 280 g/kg and the calculated requirement of arginine is (0.02 g arginine/g CP) × (Y g CP/kg diet) + (4.0 g arginine/kg diet) where Y is the dietary CP level.Abbreviations CD control diet - CP crude protein (g CP/kg diet = g nitrogen/kg diet × 6.25) - DAA dispensable amino acids - EAA essential amino acids - EAArq essential amino acid requirement     

Proline, Ornithine, Arginine and Glutamic Acid Contents in Detached Rice Leaves

The effects of water stress on the contents of proline, ornithine, arginine and glutamic acid in detached rice leaves were examined. In water stressed leaves, the content of proline was elevated to a content approximately 8-, 14- and 17-fold higher than in control leaves after 4, 8 and 12 h, respectively. We also observed that omithine and arginine contents were much higher under water stress than in control leaves. However, the content of glutamic acid in water stressed leaves was higher after 4 and 8 h and lower after 12 h than that in control leaves.     

The amnesic shellfish poison domoic acid enhances neurotoxicity by excitatory amino acids in cultured neurons

Summary A recent episode of human intoxication by cultured mussels containing a rare excitatory amino acid named domoic acid, received particular attention for its neurological implications. The intoxication produced neurological problems, such as headache, confusion, and loss of memory, particularly severe at times. Neuronal damage was found in the hippocampus and amygdala of four patients. We now report that in neuronal cultures the neurotoxicity of a domoic acid-containing mussel extract is the result of domoic acid potentiation of the excitotoxic effect of glutamic acid and aspartic acid present in high amounts in mussel tissue. Moreover, we show that subtoxic concentrations of domoic acid are sufficient to potentiate glutamic acid and aspartic acid neurotoxicity. We present evidence suggesting that the neurotoxic synergism may be due to a reduction of Mg^{+ +} block at the NMDA receptor-associated channel, following activation of NON-NMDA receptors by domoic acid.     

Glutamic Acid as a Putative Transmitter of the Interhemispheric Corticocortical Connections in the Rat

The effect of hemidecortication on the endogenous levels of amino acids in medial, sulcal, and dorsal frontal cortex as well as in parietal, temporal, and occipital cortex of the rat was investigated. Under aseptic conditions, the right cerebral cortex was aspirated by suction. Then, 21 days later, the content of glutamic acid, aspartic acid, gamma-aminobutyric acid, glycine, serine, threonine, and alanine was analyzed in six areas of the intact contralateral cortex using GLC. The results demonstrated a specific decrease in the endogenous levels of glutamic acid in both parietal and temporal cortex after hemidecortication of the contralateral side. This finding suggests that glutamic acid may serve as a neurotransmitter for some of the interhemispheric corticoparietal and corticotemporal fibers. In a follow-up experiment, the effect of a frontal lesion on the endogenous levels of the same amino acids in the striatum was also examined. In this case, the glutamic acid content exhibited a decrease of 31% relative to the control value. This observation confirms the earlier finding of a glutamate-containing pathway from the frontal cortex to the striatum.     

Cycling treatment of anaerobic and aerobic incubation increases the content of γ-aminobutyric acid in tea shoots

Summary. γ-Aminobutyric acid (GABA), a hypotensive compound, is formed from glutamic acid under anaerobic condition in tea shoots. Glutamic acid was exhausted in the first three hours of anaerobic incubation and the increase of GABA stopped. After that, when tea shoots were released under aerobic condition, glutamic acid reproduced rapidly. After one hour of aerobic incubation, tea shoots were given three hours of anaerobic incubation again and then accumulated glutamic acid changed to GABA. The content of GABA increased much more than usual anaerobic incubation. GABA was more in the tea stem than in the leaf. Received January 4, 2000 Accepted March 1, 2000     

Opossum (Didelphis virginiana) "little" and "big" gastrins

1. "Little" gastrins from most mammalian species are 17 amino acid peptides and the precursor "big" gastrins are 34 amino acid peptides. 2. "Little" gastrins of the New World hystricomorphs, guinea-pig and chinchilla, are 16 amino acid peptides due to deletion of a glutamic acid in the region 6-9 from their NH2-terminus and the corresponding "big" gastrins are 33 amino acid peptides. 3. Antral gastrins from the opossum, a New World marsupial, have a glutamic acid deletion in the same region as the hystricomorph gastrins. 4. Opossum "big" gastrin is a 33 amino acid peptide with the following sequence: less than ELGPQDLPYLTADLSKKQGPWLEEEEAYGWMDF#.     

Urinary metabolites of cis-9,10-methylene octadecanoic acid. cis-3,4-Methylene hexanedioic acid and related compounds.

cis-3,4-Methylene hexanedioic acid has been discovered in human urine. It has been isolated and identified by mass spectrometry and synthesis. The daily excretion in nine subjects on a free diet was 88 mumol/day (range, 32 to 144 mumol/day). cis-3,4-Methylene hexanedioic acid was given orally to a rat. About 90% of the dose was recovered unchanged in the urine within 24 h. Intragastric administration of cis-9,10-methylene [9,10-3H2]octadecanoic acid to rats gave four labeled urinary metabolites. The major one was cis-3,4-methylene hexanedioic acid, the others were 2,3-methylene pentanedioic acid and isomers of methylene heptanedioic acid and methylene octanedioic acid. Within 72 h, about 40% of the administered radioactivity could be recovered from the urine and another 40% from the carcass. About 20% of the recovered radioactivity was found to be water. Of the radioactivity administered to rats orally as cis-9,10-methylene [9,10-3H2]octadecanoic acid methyl ester, about 50% could be recovered from the lymph of the thoracic duct within 9 h. Intraperitoneal administration of cis-9,10-methylene octodecanoic acid methyl ester to rats gave the same metabolites. Of the given amount, 50 mol % could be recovered from the urine as cis-3,4-methylene hexanedioic acid and 19 mol % as homologues within 38 days.     

<Emphasis Type="Italic">γ</Emphasis>-Fluorinated analogues of glutamic acid and glutamine

Gamma-fluorinated analogues of glutamic acid and glutamine are compounds of biological interest. Syntheses of such compounds are extensively reviewed in this article. 4-fluoroglutamic acid was prepared as a mixture of racemic diastereomers by Michael reaction, inverse-Michael reaction or by electrophilic / nucleophilic fluorination. Optically enriched 4-fluoroglutamic acids were obtained by several resolution techniques as well as by asymmetric methodologies using the chiral pool. 4-fluoroglutamine was prepared as a mixture of stereoisomers as well as in racemic erythro and threo forms from the corresponding 4-fluoroglutamic acids using aminolysis and conventional protection and deprotection strategies. Racemic 4,4-difluoroglutamic acid was synthesized by a nitroaldol reaction and its L-enantiomer obtained via three different asymmetric routes. Racemic 4,4-difluoroglutamic acid was converted into the corresponding 4,4-difluoroglutamine using a protection / aminolysis / deprotection sequence while N-Boc-L-4,4-difluoroglutamine was prepared directly from (R)-Garner's aldehyde using a Reformatsky reaction as the key step.     

Side chain interactions in aromatic dipeptides

A study of the 100-MHZ nuclear magnetic resonace spectra in D₂O solution was made of a series of linear dipeptides of the types L -phenylalanine-L -and-D -X, and L -phenylalanine-L -and-D -Y, where X comprised a group of amino acid residues with polar side chains (X = glutamine, glutamic acid, arginine, and N^ε-acetyllysine) and Y comprised amino acid residues with purely aliphatic side chains (Y = α-aminobutyric acid and norvaline). It was found that regardless of the side chain length, resonances due to the α-methylene protons in the X and Y side chains of the L -Phe-D -Y series consistently exhibited upfield shifts greater than any other protons in these side chains, when compared to the corresponding side chain resonances of the nonaromatic dipeptide series L -Ala-L -X and L -Ala-L -Y. The magnitudes of these shielding effects were consistently and considerably greater for the L -Phe-D -X series than for the L -Phe-D -Y series. An intramolecular complex–formed by association of armatic π-electrons with the positive end of the dipole in the polar side chains—was proposed as one plausible interpretation of the enhanced shielding effects. An increase in temperature from 32 to 70–80° was sufficient to overcome the enhanced shielding attributable to the suggested complex.     

The effect of prostaglandins on the bioconversion of arachidonic acid in cervical tissue in early human pregnancy

The aim of the present study was to investigate in late first trimester and early second trimester patients whether whole cell homogenates of cervical tissue incubated with 14C-arachidonic acid was affected by pretreatment for 12 to 14 hours with PGE2 and 9-deoxo- 16,16-dimethyl-9-methylene PGE2 (9-methylene PGE2). After extraction, purification and separation, identification of the compounds found during incubation was achieved using radio-gas liquid chromatography and gas-liquid chromatography-mass spectrometry. Treatment with 9-methylene PGE2 accomplished a reduced production of 14C-labelled PGF2 alpha, -PGE2 and TxB2, while pretreatment with PGE2 induced increase in the production of 14C-6-keto-PGF1 alpha when cervical tissue homogenates were compared with specimens obtained from non-pretreated patients. Recently we reported a significantly increased formation of so far unidentified metabolite(s) in homogenates of human cervical tissue specimens obtained at or near term when compared with corresponding specimens obtained during early pregnancy. With both types of prostaglandin pretreatment there was a tendency of increased formation of these metabolites. It seems possible that the influence on the biochemistry of cervical tissue induced by PGE2 and 9-methylene PGE2 is mediated via the endogenous arachidonic acid cascade towards non-prostaglandin compound(s).     

Amino acid metabolism and the energetics of growth

The nonessential amino acids are involved in a large number of functions that are not directly associated with protein synthesis. Recent studies using a combination of transorgan balance and stable isotopic tracers have demonstrated that a substantial portion of the extra‐splanchnic flux of glutamate, glutamine, glycine and cysteine derives from tissue synthesis. A key amino acid in this respect is glutamic acid. Little glutamic acid of dietary origin escapes metabolism in the small intestinal mucosa. Furthermore, because glutamic acid is the only amino acid that can be synthesized by mammals by reductive amination of a ketoacid, it is the ultimate nitrogen donor for the synthesis of other nonessential amino acids. Because the synthesis of glutamic acid and its product glutamine involve the expenditure of adenosine triphosphate (ATP), it seems possible that nonessential amino acid synthesis might have a significant bearing on the energetics of protein synthesis and, hence, of protein deposition. This paper discusses the topic of the energy cost of protein deposition, considers the metabolic physiology of amino acid oxidation and nonessential amino acid synthesis, and attempts to combine the information to speculate on the overall impact of amino acid metabolism on the energy exchanges of animals.     

Excretion of glutamic acid in Citrobacter intermedius C3 associated with plasmid deoxyribonucleic acid.

Several mutants of Citrobacter intermedius C3 lacking both the ability to synthesize proline and the ability to excrete glutamic acid were isolated by treatment with nitrosoguanidine. No revertants for either characteristic were obtained from these mutants. The ability to excrete glutamic acid was transferred to those mutants with very high frequencies in mating experience by using auxotropic excreting strains as donors. Moreover, the ability to synthesize proline was transferred together with the ability to excrete glutamic acid when an excreting strain was used as donor. The transconjugants showed a rapid spontaneous curing of both genetic markers. It was shown by two different methods that a band of covalently closed circular deoxyribonucleic acid is present in the cesium chloride gradients corresponding to the wild type and excretor mutants. Nonexcretor mutants described herein lacked such a band. Pro + transformants that were also excretors were obtained with plasmid deoxyribonucleic acid isolated either from wild type or from an excretor mutant. These data strongly indicate that glutamic acid excretion in C. intermedius C3 is related to the presence of extrachromosomal deoxyribonucleic acid.