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Metal ion-binding ability of tetrapeptides containing alpha-aminoisobutyric acid.

alpha-Aminoisobutyric acid (Aib), one of the Calpha,alpha-disubstituted glycines, is a sterically hindered amino acid that acts as a conformational constraint in peptides. However, studies for the application of the ability of Aib to control conformation are quite few. The paper focuses on the molecular recognition ability of acyclic oligopeptides containing Aib. Liquid-liquid extraction of nine kinds of metal ions from aqueous layers to nonpolar organic layers with acyclic tetrapeptides, X-Trp-Xaa2-Gly-Xaa4-NH-Ar (X = H or C6H5CH2OCO (Z), Xaa2 = Aib or Gly, Xaa4 = Leu or Ala, Ar = phenyl or 3,5-dimethylphenyl) was examined using picrate as the anion of ion pairs. The extraction behaviour of the metal ions with the tetrapeptides was investigated in the pH range from 3 to 9. In the case of basic pH regions, Cu(II) and Ag(I) were effectively extracted with Trp-Aib-Gly-Leu-NH-Ar. Pd(II) was specifically extracted with Trp-Aib-Gly-Leu-NH-Ar in acidic pH regions. The extraction percent (%E) of the peptide host, which has a 3,5-dimethylphenyl group, was even larger than that of the host, which has a phenyl group. Moreover, Pd(II) was extracted with a peptide host which has Leu and a 3,5-dimethylphenyl group in the absence of picrate as the anion of ion pairs. The free alpha-amino group, the turn conformation and the hydrophobicity of peptide molecules were important factors for the extraction of the metals.

Aminoisobutyric Acids↗

Steady-state distribution of cycloleucine and alpha-aminoisobutyric acid between plasma and cerebrospinal fluid.

Estimates of the steady-state distribution ratios of two nonmetabolizable amino acids, alpha-aminoisobutyric acid and aminocyclopentane carboxylic acid (cycloleucine), between plasma and cerebrospinal fluid were made with a view to establishing whether or not the low values found with metabolizable amino acids, such as glycine or leucine, could be accounted for by uptake and metabolism by the brain. The estimates, based on the ratios found after i.p. injections either in bolus form or by implantation of "osmotic pumps" containing the labeled amino acids, were comparable with those found for metabolizable amino acids.

Amino Acids↗

Inhibition of alpha-aminoisobutyric acid uptake by diisothiocyanostilbenedisulphonic acid in the amphibian cornea.

alpha-Aminoisobutyric acid accumulation of the toad's (Bufo marinus) cornea and lens is inhibited by 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid. This effect is seen at pH 8.4; at pH 7.4 a small increase in aminoisobutyric acid uptake was observed. Efflux of aminoisobutyric acid is unchanged by diisothiocyanostilbenedisulphonic acid at either pH. The inhibitory effect of diisothiocyanostilbenedisulphonic acid on aminoisobutyric acid accumulation appears to reflect a direct action on membrane mechanisms that mediate its influx.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Influx of gamma-aminobutyric acid and alpha-aminoisobutyric acid into mouse cerebrum slices incubated in a pyruvate medium with or without added glucose or glucose analogues compared with influx from a glucose medium.

Concentrative influx of gamma-aminobutyric acid (GABA) and alpha-amino-isobutyric acid (AIB) into incubated mouse cerebrum slices is decreased when pyruvate is substituted for glucose. Influx of GABA from pyruvate medium is not increased by presence of glucose, 2-deoxy-D-glucose (2-DOG), or 3-O-methyl-D-glucose (3-O-MeG). Influx of AIB is restored to the rate from glucose medium if 2-DOG is present initially, but is not restored if 2-DOG is added with AIB. Influx is not restored if 3-O-MeG is present initially, but is restored if 3-O-MeG is added with AIB. Influx is restored if glucose is present initially or is added with AIB.

Aminoisobutyric Acids↗

Occurrence of beta-aminoisobutyric acid in Mytilus edulis.

beta-Aminoisobutyric acid was isolated from organ extracts of Mytilus edulis. Ion-exchange resins and large-scale paper chromatography were used to isolate minute quantities of the compound. beta-Aminoisobutyric acid was identified by paper chromatography in several solvents and by conversion to DNP-beta-aminoisobutyric acid and subsequent chromatography of the derivative in several solvents.

Amino Acids↗

Elevation of serum levels of beta-aminoisobutyric acid in uremic patients and the toxicity of the amino acid.

A reliable method for the determination of beta-aminoisobutyric acid in serum was developed utilizing an automated amino acid analyzer. The serum concentrations of beta-aminoisobutyric acid were determined in 20 normal subjects and in 71 uremic patients. The mean serum level of beta-aminoisobutyric acid was markedly increased in the uremic patients to 0.856 +/- 0.910 (mean +/- SD) mg/100 ml as compared with a normal value of 0.026 +/- 0.027 mg/100 ml. The distribution of serum beta-aminoisobutyric acid level in uremic patients was wide-spread, and there was no correlation between the serum levels of the amino acid and those of urea nitrogen, creatinine and uric acid. The toxicity of beta-aminoisobutyric acid on mice with acute renal failure induced by uranyl acetate was investigated and compared with that of alpha-amino-n-butyric acid and gamma-amino-n-butyric acid. All mice given more than 4 g/kg body wt of beta-aminoisobutyric acid showed twitching and cramps, and some of them died. However, the control mice given an equivalent dose of alpha-amino-n-butyric acid or gamma-amino-n-butyric acid showed no change. These results suggest that beta-aminoisobutyric acid might be a factor influencing the development and progression of uremic toxemia.

Acute Kidney Injury↗

Energetics of sodium-dependent alpha-aminoisobutyric acid transport in the moderate halophile Vibrio costicola.

The energetics of alpha-aminoisobutyric acid transport were examined in Vibrio costicola grown in a medium containing the NaCl content (1 M) optimal for growth. Respiration rate, the membrane potential (delta psi) and alpha-aminoisobutyric acid transport had similar pH profiles, with optima at 8.5-9.0. Cells specifically required Na+ ions to transport alpha-aminoisobutyric acid and to maintain the highest delta psi (150-160 mV). Sodium was not required to sustain high rates of O2-uptake. Delta psi (and alpha-aminoisobutyric acid transport) recovered fully upon addition of Na+ to Na+-deficient cells, showing that Na+ is required in formation or maintenance of the transmembrane gradients of ions. Inhibitions by protonophores, monensin, nigericin and respiratory inhibitors revealed a close correlation between the magnitudes of delta psi and alpha-aminoisobutyric acid transport. Also, dissipation of delta psi with triphenylmethylphosphonium cation abolished alpha-aminoisobutyric acid transport without affecting respiration greatly. On the other hand, alcohols which stimulated respiration showed corresponding increases in alpha-aminoisobutyric acid transport, without affecting delta psi. Similarly, N,N'-dicyclohexylcarbodiimide (10 microM) stimulated respiration and alpha-aminoisobutyric acid transport and did ot affect delta psi, but caused a dramatic decline in intracellular ATP content. From these, and results obtained with artificially established energy sources (delta psi and Na+ chemical potential), we conclude that delta psi is obligatory for alpha-aminoisobutyric acid transport, and that for maximum rates of transport an Na+ gradient is also required.

Aminoisobutyric Acids↗

Evidence for the release of newly acquired ascorbate and alpha-aminoisobutyric acid from the cytosol of adrenomedullary chromaffin cells through specific transporter mechanisms.

Primary cultures of bovine adrenomedullary cells actively take up ascorbic acid and alpha-aminoisobutyric acid (AIB). Following a brief incubation with L-[14C] ascorbic acid and alpha-[methyl-3H]aminoisobutyric acid, cells stimulated with the nicotinic agonist 1,1-dimethyl-4-phenylpiperazinium iodide or by membrane depolarization with high [K+] or veratridine released newly acquired ascorbic acid (NA-ascorbate) and AIB. NA-ascorbate and endogenous catecholamines are differentially released under a variety of conditions suggesting that release of both substances cannot originate from the same subcellular compartment. In contrast, the release profile for NA-ascorbate and AIB, a putative cytosolic marker, suggest that both of these molecules are released from a cytosolic compartment. Cells permeabilized with the detergent digitonin release catecholamines only in the presence of external Ca2+, whereas release of NA-ascorbate and AIB is Ca2+-independent and time- and detergent concentration-dependent. If the osmolality of the external medium is made either hyper- or hypoosmotic, 1,1-dimethyl-4-phenylpiperazinium iodide-induced release of endogenous catecholamines is inhibited. Release of NA-ascorbate and AIB, however, is progressively inhibited with increasing osmolality and enhanced with decreasing osmolality. Furthermore, differential release of NA-ascorbate and AIB as compared to soluble acetylcholinesterase, which is apparently released form the cisternae of the endoplasmic reticulum, was also observed. To determine the mechanism by which NA-ascorbate and AIB are released from the cell, the requirements for their maximal release were investigated. Release of NA-ascorbate and AIB was sensitive to inhibitors (both metabolic and transport) and to changes in the external ionic environment. The metabolic inhibitors carbonyl cyanide p-trifluoromethoxyphenylhydrazone and KCN (when incubated simultaneously with 2-deoxyglucose) inhibited NA-ascorbate and AIB release by greater than 75%. In contrast, the Na+-K+-ATPase inhibitor ouabain enhanced veratridine-induced release of NA-ascorbate by nearly 100% and had an even greater effect on AIB release. Changes in the external ionic environment (i.e. Na+ and/or Cl- substitution) inhibited both NA-ascorbate and AIB release to varying degrees. Substitution of Cl- by various anions inhibited NA-ascorbate and AIB release to a much greater degree than endogenous catecholamine release. Complete substitution of NaCl with sucrose inhibited release of NA-ascorbate and AIB release by greater than 80%, while Na+ substituted with Li+ inhibited release of all three molecules by about 50%.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholinesterase↗

Effect of dietary n-3 fatty acid deficiency on blood-to-brain transfer of sucrose, alpha-aminoisobutyric acid and phenylalanine in the rat.

Possible alterations in blood-to-brain unidirectional transport of sucrose (mol. wt., 342), alpha-aminoisobutyric acid (mol. wt., 104), and L-phenylalanine (mol. wt., 165) induced by a diet deficient in n-3 polyunsaturated fatty acids were studied with respect to blood-brain barrier function. Two groups of rats were for to two generations with a semisynthetic diet. One group of rats was fed a peanut oil+rapeseed oil diet which contained both essential fatty acids: linoleic acid (18:2 n-6) and alpha-linolenic acid, (18:3 n-3). Another group was fed a diet of peanut oil, this diet (containing 18:2 n-6) was deficient in alpha-linolenic acid. The experiments were performed at 6 months of age. Unidirectional transfer rate constants (Ki) of sucrose, alpha-aminoisobutyric acid and L-phenylalanine were measured. The diet based on peanut oil (deficient in n-3) caused a greater blood-to-brain transport of sucrose but not of alpha-aminoisobutyric acid or L-phenylalanine. These observations indicate that regardless of the mechanisms involved, alterations in essential fatty acids induced by diet can modulate to some extent the blood-brain transport of hydrophilic molecules without a carrier.

Aminoisobutyric Acids↗

Synthesis of [11C-methyl]-alpha-aminoisobutyric acid (AIB).

Alpha-aminoisobutyric acid (AIB) labeled with the cyclotron-produced, positron-emitting radionuclide 11C has been synthesized with the label in the alpha-methyl group. Our previously published synthesis of [11C] AIB in the carboxyl position from [11C] HCN requires a rigorous quality assurance program to ensure that the concentration of cyanide in the final product is below certain levels. This can be avoided using the method described here with [11C] CH3I. The radiochemical yield calculated to end of bombardment (EOB) was 45-55% from [11C] CO2 with radiochemical purity of [11C-methyl] AIB exceeding 99%. Synthesis times from [11C] CO2 were about 55 min. Specific activities of 1 Ci/mumol were achieved on average. It has been shown that [11C-carboxyl] AIB is a useful imaging agent in patients with soft tissue cancers and melanoma, and it demonstrates tumor uptake in a spectrum of other animal tumor models. Because AIB is a nonmetabolized amino acid, [11C-methyl] AIB should be equally as useful. Either agent can be employed for the quantification of the A-type amino acid transport system in vivo with PET.

Aminoisobutyric Acids↗

Amino acid uptake in the developing chick embryo heart. The effect of insulin on alpha-aminoisobutyric acid accumulation.

1. The uptake of (14)C-labelled alpha-aminoisobutyric acid by 5-day-old chick embryo hearts was investigated in vitro, together with the effect of insulin thereon. 2. At equilibrium the distribution ratio of this amino acid analogue between intracellular and extracellular water attained values greater than unity. Insulin enhanced the rate of alpha-aminoisobutyric acid accumulation and increased the value of its final concentration in the cell water. 3. The rate of alpha-aminoisobutyric acid accumulation and the effect of insulin on it were independent of the presence of glucose in the incubation medium. Bovine and chicken insulin were equally effective, and the action of the hormone was specifically prevented by an anti-insulin serum but not by puromycin. 4. A linear relationship was observed between the intracellular accumulation of the analogue and the logarithm of the insulin concentration in the range 50muunits-100m-units/ml. of incubation medium. 5. Evidence was obtained for the occurrence of two different transport processes for alpha-aminoisobutyric acid in the chick embryo heart: one subject to saturation and one that was not saturated by reasonable concentrations of the analogue. Insulin increased the effectiveness of the saturable component, increasing the maximal velocity of transport without altering the concentration for half-maximal velocity of transport, and decreased the contribution of the non-saturable component.

Amino Acids↗

Imaging studies of patients with malignant fibrous histiocytoma using C-11-alpha-aminoisobutyric acid (AIB).

Alpha-aminoisobutyric acid (AIB), a synthetic, nonmetabolized amino acid which is rapidly transported into viable cells by the A-type or alanine-preferring amino acid transport system, has been labeled with the short-lived, positron-emitting radionuclide carbon-11. Carbon-11 labeled AIB is currently being evaluated as a tumor imaging agent for in vivo amino acid transport studies in patients with cancer. In this study, C-11 AIB was used to image two patients with malignant fibrous histiocytoma (MFH), a pleomorphic sarcoma. Following intravenous administration of C-11 AIB, tumors in the distal femur of one patient and in the anterior chest wall of another patient were well visualized using high energy gamma scintigraphy. Since therapy may alter the accumulation of amino acids in tumor tissue, studies using C-11 AIB in patients with MFH before and after chemotherapy are in progress.

Adult↗