Kegg Pathway: Valine, leucine and isoleucine degradation

Biochim Biophys Acta. 2008 Oct; 1780(10): 1115-1120
Herningtyas EH, Okimura Y, Handayaningsih AE, Yamamoto D, Maki T, Iida K, Takahashi Y, Kaji H, Chihara K

The effect of amino acid on muscle protein degradation remains unclear. Recent studies have elucidated that proteolysis in catabolic conditions occurs through ubiquitin-proteasome proteolysis pathway and that muscle-specific ubiquitin ligases (atrogin-1 and MuRF1) play an important role in protein degradation. In the present study, we examined the direct effect of 5 mM amino acids (leucine, isoleucine, Valine, glutamine and arginine) on atrogin-1 and MuRF1 levels in C2C12 muscle cells and the involved intracellular signal transduction pathway. leucine, isoleucine and valine suppressed atrogin-1 and MuRF1 mRNA levels ( approximately 50%) at 6 and 24 h stimulations. Arginine showed a similar effect except at 24 h-treatment for atrogin-1 mRNA. However, glutamine failed to reduce atrogin-1 and MuRF1 mRNA levels. The inhibitory effect of leucine, isoleucine or arginine on atrogin-1 mRNA level was reversed by rapamycin, although wortmannin did not reverse the effect. PD98059 and HA89 reduced basal atrogin-1 level without influencing the inhibitory effects of those amino acids. The inhibitory effect of leucine, isoleucine or arginine on MuRF1 mRNA levels was not reversed by rapamycin. Taken together, these findings indicated that leucine, isoleucine and arginine decreased atrogin-1 mRNA levels via mTOR and that different pathways were involved in the effect of those amino acids on MuRF1 mRNA levels.

Magn Reson Imaging. 2008 Jul 2;
Balasubramanian K, Kumar S, Singh RR, Sharma U, Ahuja V, Makharia GK, Jagannathan NR

Metabolism of the colonic mucosa of patients with ulcerative colitis (UC; n=31) and Crohn's disease (CD; n=26) and normal mucosa (control, n=26) was investigated using in vitro high-resolution proton magnetic resonance spectroscopy. Of the 31 UC patients, 20 were in the active phase and 11 were in the remission phase of the disease. Out of 26 CD patients, 20 were in the active phase, while 6 were in the remission phase of the disease. Twenty-nine metabolites were assigned unambiguously in the perchloric acid extract of colonic mucosa. In the active phase of UC and CD, significantly lower (Pleucine, leucine, Valine, alanine, glutamate and glutamine), membrane components (choline, glycerophosphorylcholine and myo-inositol), lactate and succinate were observed compared to normal mucosa of controls. Patients in the active phase of UC and CD also showed increased level of alpha-glucose compared to normal mucosa. Altered level of metabolites indicates decreased protein and carbohydrate metabolism, thereby decreased energy status and deterioration of mucosa integrity during chronic inflammation. In the remission phase of UC and CD, the concentration of most of the metabolites was similar to controls except for lower values of lactate, glycerophosphorylcholine and myo-inositol in UC and Lac in CD. Formate was significantly lower in patients with the active phase of UC compared to patients with the active phase of CD, suggesting the potential of in vitro MRS in the differentiation of these two diseases.

J Appl Physiol. 2008 Aug; 105(2): 643-51
Galloway SD, Talanian JL, Shoveller AK, Heigenhauser GJ, Spriet LL

This study examined 1) the plasma taurine response to acute oral taurine supplementation (T), and 2) the effects of 7 days of T on muscle amino acid content and substrate metabolism during 2 h of cycling at approximately 60% peak oxygen consumption (Vo(2peak)). In the first part of the study, after an overnight fast, 7 volunteers (28 +/- 3 yr, 184 +/- 2 cm, 88.0 +/- 6.6 kg) ingested 1.66 g oral taurine doses with breakfast (8 AM) and lunch (12 noon), and blood samples were taken throughout the day. In the second part of the study, eight men (22 +/- 1 yr, 181 +/- 1 cm, 80.9 +/- 3.8 kg, 4.21 +/- 0.16 l/min Vo(2peak)) cycled for 2 h after 7 days of placebo (P) ingestion (6 g glucose/day) and again following 7 days of T (5 g/day). In the first part of the study, plasma taurine was 64 +/- 4 muM before T and rose rapidly to 778 +/- 139 muM by 10 AM and remained elevated at noon (359 +/- 56 muM). Plasma taurine reached 973 +/- 181 muM at 1 PM and was 161 +/- 31 muM at 4 PM. In the second part of the study, seven days of T had no effect on muscle taurine content (mmol/kg dry muscle) at rest (P, 44 +/- 15 vs. T, 42 +/- 15) or after exercise (P, 43 +/- 12 vs. T, 43 +/- 11). There was no difference in muscle glycogen or other muscle metabolites between conditions, but there were notable interaction effects for muscle Valine, isoleucine, leucine, cystine, glutamate, alanine, and arginine amino acid content following exercise after T. These data indicate that 1) acute T produces a 13-fold increase in plasma taurine concentration; 2) despite the ability to significantly elevate plasma taurine for extended periods throughout the day, 7 days of T does not alter skeletal muscle taurine content or carbohydrate and fat oxidation during exercise; and 3) T appears to have some impact on muscle amino acid response to exercise.

Br Poult Sci. 2008 May; 49(3): 299-307
Imanari M, Kadowaki M, Fujimura S

1. The effects of dietary branched-chain amino acids (BCAAs) including leucine (Leu), isoleucine (Ile) and valine (Val) on taste-active components, especially free glutamate (Glu), in meat were investigated. 2. Broiler chickens (28 d old) were given varied dietary BCAA levels for 10 d before marketing. Dietary BCAA content ratios were either 100:100:100 (Low Leu group), 150:100:100 (Control group) or 150:150:150 (High Ile + Val group) for Leu:Ile:Val (% of each BCAA requirement according to NRC, 1994). Taste-related components of meat (free amino acids and ATP metabolites) and sensory scores of meat soup were estimated. 3. Free Glu content, the main taste-active component of meat, was significantly increased by dietary BCAA. Compared to the Control group, free Glu content increased by 30% in the High Ile + Val group. However, the inosine monophosphate (IMP) content in meat did not change among groups. 4. Sensory evaluation of meat soups showed that Control and High Ile + Val groups had different meat flavours. The sensory score of overall taste intensity was significantly higher in the High Ile + Val group. 5. These results suggest that dietary BCAA concentrations regulate free Glu in meat. Increasing dietary Ile + Val induces an increase in free Glu content of meat, improves meat taste and is more effective for increasing free Glu content in meat than decreasing dietary Leu level.

J Bacteriol. 2008 Aug; 190(15): 5291-9
Lemos JA, Nascimento MM, Lin VK, Abranches J, Burne RA

The RelA, RelP, and RelQ enzymes are responsible for the production of the alarmone (p)ppGpp in Streptococcus mutans. A strain lacking all three synthetases (DeltarelAPQ) does not grow in minimal medium lacking the branched-chain amino acids (BCAA) leucine or valine but grows well if isoleucine is also omitted. Here, we investigated whether there was a correlation between growth in the absence of leucine and valine with (p)ppGpp pools and the activation of CodY. By using a combination of single, double, and triple mutants lacking the (p)ppGpp synthetase enzymes, we demonstrated that the ability to grow in the absence of leucine or valine required basal levels of (p)ppGpp production by RelP and RelQ. The introduction of a codY mutation into the DeltarelAPQ strain fully restored growth in medium lacking leucine or Valine, revealing that the growth-defective phenotype of DeltarelAPQ was directly linked to CodY. Lowering GTP levels through the addition of decoyinine did not alleviate CodY repression or affect the expression of genes involved in BCAA biosynthesis, suggesting that S. mutans CodY is not activated by GTP. The results of phenotypic studies revealed that the codY mutant had a reduced capacity to form biofilms and that its growth was more sensitive to low pH, showing a role for CodY in two key virulence properties of S. mutans. Microarray results revealed the extent of the CodY regulon. Notably, the identification of putative CodY-binding boxes upstream of genes that were downregulated in the codY mutant indicates that CodY may also function as a transcriptional activator in S. mutans.

Pediatr Int. 2008 Jun; 50(3): 312-4
Silao CL, Padilla CD, Matsuo M

BACKGROUND: Maple syrup urine disease (MSUD) is an autosomal recessive disorder caused by defective activity of the branched-chain alpha-ketoacid dehydrogenase enzyme complex. Early diagnosis and management of MSUD are imperative for preventing permanent neurological impairments. In the Philippines, a 4.7 kb deletion in the dihydrolipoamide branched-chain transacylase E2 (DBT) gene has been commonly identified in MSUD patients. Polymerase chain reaction (PCR) amplification of a junction fragment between intron 10 and exon 11 has been used to detect this deletion. The purpose of the present paper was to use PCR-based mutation detection of the deletion mutation to diagnose MSUD in neonates in order to provide proper diagnosis and effective treatment. METHODS: A region encompassing exon 11 and the junction fragment of the E2 (DBT) gene was PCR amplified from genomic DNA prepared from two neonates at risk for MSUD. RESULTS: PCR amplification of both exon 11 and the junction fragment from one of the neonates demonstrated that this case was a heterozygous carrier of the deletion. Thus, normal feeding was started. For the other neonate, PCR amplification of the junction fragment was successful, whereas the region encompassing exon 11 was not amplified. This neonate was genotyped as homozygous for the deletion, and treatment for MSUD was provided immediately. CONCLUSION: Examination of the deletion mutation in the E2 (DBT) gene facilitated early MSUD diagnosis and was beneficial for the determination of the proper course of treatment.

J Forensic Sci. 2008 May; 53(3): 730-3
Girela E, Villanueva E, Irigoyen P, Girela V, Hernández-Cueto C, Peinado JM

We studied free amino acids in vitreous humor and cerebrospinal fluid from 58 cadavers in the course of routine medicolegal autopsies in the city of Granada. The main objective was to establish whether free amino acids contents in these fluids were related with the cause of death, postmortem interval, and severity of the classic signs of asphyxia. The amino acids (aspartic acid, glutamic acid, serine, glutamine, glycine/threonine/histidine, citruline, arginine, alanine, taurine, GABA, tirosine, Valine, methionine, isoleucine, phenylalanine/tryptophan, leucine, and lysine) were quantified by high performance liquid chromatography. There were no statistically significant differences in amino acids concentrations in vitreous humor when the different causes of death were considered. Our results did not show any statistically significant relationship when asphyxial score was plotted against the vitreous content of each amino acid. A statistically significant increase with postmortem interval was observed in vitreous taurine (r = 0.3191, p = 0.01461), glutamate (r = 0.4323, p = 0.0007) and particularly in aspartate (r = 0.4508, p = 0.0003).

Am J Vet Res. 2008 May; 69(5): 617-24
Frantz NZ, Andrews GA, Tokach MD, Nelssen JL, Goodband RD, Derouchey JM, Dritz SS

OBJECTIVE: To evaluate dietary ingredients involved in cartilage and bone metabolism and their influence on osteochondrosis lesions in swine. ANIMALS: 80 crossbred gilts (mean initial weight, 39 kg). PROCEDURES: Pigs (10 pigs/treatment) were fed a corn-soybean meal basal (control) diet or the basal diet supplemented with additional minerals (copper and manganese or silicon), amino acids (proline and glycine; a combination of leucine, isoleucine, and valine; or methionine and threonine), or fatty acids (provided by fish oil) for 84 days. Pigs were then slaughtered and the distal portion of the left femur was collected for determination of osteochondrosis lesions at the femoral condyle. After evaluation of external joint surfaces, the distal portion of the femur was sectioned to evaluate lesions in the growth plate and articular cartilage. Additionally, a cartilage specimen was obtained from the patella for analysis. RESULTS: Pigs fed diets containing high amounts of methionine and threonine or the diet containing all additional ingredients had significantly lower total severity scores, compared with scores for pigs fed the control diet or a diet supplemented with fish oil. Pigs fed diets containing additional proline and glycine, copper and manganese, methionine and threonine, or all additional ingredients had significantly lower overall scores, compared with scores for pigs fed the control diet or a diet supplemented with fish oil. CONCLUSIONS AND CLINICAL RELEVANCE: Dietary manipulation decreased the severity of osteochondrosis lesions, compared with results for pigs fed a control diet. However, additional research on optimal concentrations and combinations of dietary components is needed.

FEBS J. 2008 Jun; 275(11): 2727-37
Numa N, Ishida Y, Nasu M, Sohda M, Misumi Y, Noda T, Oda K

Hypophosphatasia, a congenital metabolic disease related to the tissue-nonspecific alkaline phosphatase gene (TNSALP), is characterized by reduced serum alkaline phosphatase levels and defective mineralization of hard tissues. A replacement of valine with alanine at position 406, located in the crown domain of TNSALP, was reported in a perinatal form of hypophosphatasia. To understand the molecular defect of the TNSALP (V406A) molecule, we examined this missense mutant protein in transiently transfected COS-1 cells and in stable CHO-K1 Tet-On cells. Compared with the wild-type enzyme, the mutant protein showed a markedly reduced alkaline phosphatase activity. This was not the result of defective transport and resultant degradation of TNSALP (V406A) in the endoplasmic reticulum, as the majority of newly synthesized TNSALP (V406A) was conveyed to the Golgi apparatus and incorporated into a cold detergent insoluble fraction (raft) at a rate similar to that of the wild-type TNSALP. TNSALP (V406A) consisted of a dimer, as judged by sucrose gradient centrifugation, suggestive of its proper folding and correct assembly, although this mutant showed increased susceptibility to digestion by trypsin or proteinase K. When purified as a glycosylphosphatidylinositol-anchorless soluble form, the mutant protein exhibited a remarkably lower Kcat/Km value compared with that of the wild-type TNSALP. Interestingly, leucine and isoleucine, but not phenylalanine, were able to substitute for Valine, pointing to the indispensable role of residues with a longer aliphatic side chain at position 406 of TNSALP. Taken together, this particular mutation highlights the structural importance of the crown domain with respect to the catalytic function of TNSALP.

Plant Physiol. 2008 Jun; 147(2): 912-21
Gattolin S, Newbury HJ, Bale JS, Tseng HM, Barrett DA, Pritchard J

We have used high-sensitivity capillary electrophoresis coupled to a laser-induced fluorescence detection method to quantify 16 amino acids in wheat (Triticum aestivum) sieve tube (ST) samples as small as 2 nL collected by severing the stylets of feeding aphids. The sensitivity of the method was sufficient to determine a quantitative amino acid profile of individual STs without the need to bulk samples to produce larger volumes for analysis. This allowed the observation of the full range of variation that exists in individual STs. Some of the total concentrations of amino acids recorded are higher than those reported previously. The results obtained show variation in the concentrations of phenylalanine (Phe), histidine/valine (His/Val), leucine/isoleucine (Leu/Ile), arginine, asparagine, glutamine, tyrosine (Tyr), and lysine (Lys) across the ST samples. These could not be explained by plant-to-plant variation. Statistical analyses revealed five analytes (Tyr, Lys, Phe, His/Val, and Leu/Ile) that showed striking covariation in their concentrations across ST samples. A regression analysis revealed a significant relationship between the concentrations of Tyr, Lys, Phe, Leu/Ile, His/Val, asparagine, arginine, and proline and the time of collection of ST samples, with these amino acids increasing in concentration during the afternoon. This increase was confirmed to occur in individual STs by analyzing samples obtained from stylet bundles exuding for many hours. Finally, an apparent relationship between the exudation rate of ST sap and its total amino acid concentration was observed: samples containing higher total amino acid concentrations were observed to exude from the severed stylet bundles more slowly.

J Alzheimers Dis. 2008 Mar; 13(2): 123-35
Willis M, Prokesch M, Hutter-Paier B, Windisch M, Stridsberg M, Mahata SK, Kirchmair R, Wietzorrek G, Knaus HG, Jellinger K, Humpel C, Marksteiner J

Chromogranin B and secretogranin II are major soluble constituents of large dense core vesicles of presynaptic structures and have been found in neuritic plaques of Alzheimer patients. We examined the distribution and expression of these peptides in both transgenic mice over expressing human amyloid-beta protein precursor APP751 with the London (V717I) and Swedish (K670M/N671L) mutations and in human post-mortem brain. In transgenic mice, the number of amyloid-beta plaques and chromogranin immunopositive plaques increased from 6 to 12 months. About 60% of amyloid-beta plaques were associated with chromogranin B and about 40% with secretogranin II. Chromogranin immunoreactivity appeared mainly as swollen dystrophic neurites. Neither synaptophysin- nor glial fibrillary acidic protein- immunoreactivity was expressed in chromogranin immunoreactive structures at any timepoint. Density of chromogranin peptides in hippocampal structures did not change in transgenic animals at any timepoint, even though animals had a poorer performance in the Morris water maze task. In conclusion, our findings in transgenic animals partly resembled findings in Alzheimer patients. Chromogranin peptides were associated with amyloid-beta plaques, but were not reduced in specific brain areas as previously reported by our group. Therefore specific changes of chromogranin peptides observed in Alzheimer patients can be related to amyloid-beta pathology only.

Pediatrics. 2008 Apr; 121(4): e920-6
Scholl-Bürgi S, Haberlandt E, Heinz-Erian P, Deisenhammer F, Albrecht U, Sigl SB, Rauchenzauner M, Ulmer H, Karall D

OBJECTIVE: The purpose of this work was to investigate the influence of age, gender, and antiepileptic therapy on amino acid cerebrospinal fluid/plasma ratios in children. PATIENTS AND METHODS: Concentrations of 17 amino acids measured by ion-exchange chromatography with ninhydrin detection in plasma and cerebrospinal fluid from 68 patients with neurologic diseases were used to calculate their cerebrospinal fluid/plasma ratios (70 measurements; 28 female patients [29 punctures] and 40 male patients [41 punctures]). Age dependence and the effects of gender and antiepileptic medication on amino acid cerebrospinal fluid/plasma ratios were investigated by linear multiple regression analysis, and nonstandardized predicted mean values for 2 age groups were calculated (cutoff: 3 years old). RESULTS: The cerebrospinal fluid/plasma ratios ranged between 0.02 for glycine and 0.93 for glutamine. Age had a significant influence on cerebrospinal fluid/plasma ratios for Valine, isoleucine, leucine, and tyrosine, with higher ratios in younger children. Gender had a significant influence only on the glutamine cerebrospinal fluid/plasma ratio (female patients had lower ratios). Cerebrospinal fluid/plasma ratios of glutamine and tyrosine were significantly elevated by valproate therapy and those of serine, asparagine, glutamine, Valine, methionine, and phenylalanine by phenobarbital therapy. No significant influence of age, gender, and antiepileptic drugs was detectable on cerebrospinal fluid/plasma ratios of threonine, proline, glycine, alanine, histidine, ornithine, lysine, and arginine. CONCLUSIONS: Cerebrospinal fluid/plasma ratios, especially for essential neutral amino acids and for serine, asparagine, and glutamine were influenced to different degrees by age, gender, and antiepileptic therapy.

J Neurochem. 2008 May; 105(4): 1176-86
Murín R, Schaer A, Kowtharapu BS, Verleysdonk S, Hamprecht B

The branched-chain amino acids (BCAAs)--isoleucine, leucine, and valine--belong to the limited group of substances transported through the blood-brain barrier. One of the functions they are thought to have in brain is to serve as substrates for meeting parenchymal energy demands. Previous studies have shown the ubiquitous expression of a branched-chain alpha-keto acid dehydrogenase among neural cells. This enzyme catalyzes the initial and rate-limiting step in the irreversible degradative pathway for the carbon skeleton of valine and the other two branched-chain amino acids. Unlike the acyl-CoA derivates in the irreversible part of valine catabolism, 3-hydroxyisobutyrate could be expected to be released from cells by transport across the mitochondrial and plasma membranes. This could indeed be demonstrated for cultured astroglial cells. Therefore, to assess the ability of neural cells to make use of this valine-derived carbon skeleton as a metabolic substrate for the generation of energy, we investigated the expression in cultured neural cells of the enzyme processing this hydroxy acid, 3-hydroxyisobutyrate dehydrogenase (HIBDH). To achieve this, HIBDH was purified from bovine liver to serve as antigen for the production of an antiserum. Affinity-purified antibodies against HIBDH specifically recognized the enzyme in liver and brain homogenates. Immunocytochemistry demonstrated the ubiquitous expression of HIBDH among cultured glial (astroglial, oligodendroglial, microglial, and ependymal cells) and neuronal cells. Using an RT-PCR technique, these findings were corroborated by the detection of HIBDH mRNA in these cells. Furthermore, immunofluorescence double-labeling of astroglial cells with antisera against HIBDH and the mitochondrial marker pyruvate dehydrogenase localized HIBDH to mitochondria. The expression of HIBDH in neural cells demonstrates their potential to utilize valine imported into the brain for the generation of energy.

J Ren Nutr. 2008 Mar; 18(2): 239-47
Małgorzewicz S, Debska-Slizień A, Rutkowski B, Lysiak-Szydłowska W

OBJECTIVE: The aim of this study was to evaluate the correlation between the serum concentration of amino acids (AAs) and nutritional status in hemodialysis (HD) patients. METHODS: This study was performed in 22 HD patients dialyzed for 10 to 288 months, and in a control group of 20 healthy volunteers. Nutritional status was determined by the subjective global assessment method and by measuring albumin concentration. Body composition was determined using the parameters of body mass index, and the percentage of body fat and lean body mass (as measured by the near-infrared method). We measured C-reactive protein (CRP) as a marker of inflammatory status. Serum concentrations of 20 AAs were measured by precolumn orthophtalaldehyde derivatization, applying high-performance liquid chromatography (Hitachi-Merck HPLC, Tokyo, Japan) equipped with a C-18 reversed-phase column and a methanol/acetate buffer gradient. RESULTS: Thirteen of 22 (59%) patients were of good nutritional status, and 9/22 (41%) were malnourished, including 1 person with severe malnutrition. In dialyzed patients compared with control subjects, a decreased concentration of essential and nonessential AAs was observed (P < .05). Concentrations of the majority of studied AAs (16 out of 20) were lower in patients dialyzed for a period >2 years, compared with patients dialyzed for a shorter time. The ratio of branched-chain amino acids (BCAAs) to aromatic AAs was lower in the dialyzed group compared with control subjects. This ratio was also lower in patients dialyzed longer compared with patients dialyzed for <2 years. No correlation between the concentration of some AAs and CRP level or dialysis adequacy was observed. In the malnourished group, an insignificantly lower concentration of some essential AAs (lysine, leucine, isoleucine, Valine, and threonine), and a significantly higher (P = .04) concentration of CRP, were observed. CONCLUSION: Despite quite good nutritional status, dialyzed patients present abnormalities in their AA profiles. Moreover, a significant decrease of BCAA concentration is related to calorie-protein malnutrition, inflammation, and a long period of hemodialysis.

J Chem Phys. 2008 Feb 7; 128(5): 052314
Helmus JJ, Nadaud PS, Höfer N, Jaroniec CP

We describe three- and four-dimensional semiconstant-time transferred echo double resonance (SCT-TEDOR) magic-angle spinning solid-state nuclear magnetic resonance (NMR) experiments for the simultaneous measurement of multiple long-range (15)N-(13)C(methyl) dipolar couplings in uniformly (13)C, (15)N-enriched peptides and proteins with high resolution and sensitivity. The methods take advantage of (13)C spin topologies characteristic of the side-chain methyl groups in amino acids alanine, isoleucine, leucine, methionine, threonine, and valine to encode up to three distinct frequencies ((15)N-(13)C(methyl) dipolar coupling, (15)N chemical shift, and (13)C(methyl) chemical shift) within a single SCT evolution period of initial duration approximately 1(1)J(CC) (where (1)J(CC) approximately 35 Hz, is the one-bond (13)C(methyl)-(13)C J-coupling) while concurrently suppressing the modulation of NMR coherences due to (13)C-(13)C and (15)N-(13)C J-couplings and transverse relaxation. The SCT-TEDOR schemes offer several important advantages over previous methods of this type. First, significant (approximately twofold to threefold) gains in experimental sensitivity can be realized for weak (15)N-(13)C(methyl) dipolar couplings (corresponding to structurally interesting, approximately 3.5 A or longer, distances) and typical (13)C(methyl) transverse relaxation rates. Second, the entire SCT evolution period can be used for (13)C(methyl) and/or (15)N frequency encoding, leading to increased spectral resolution with minimal additional coherence decay. Third, the experiments are inherently "methyl selective," which results in simplified NMR spectra and obviates the use of frequency-selective pulses or other spectral filtering techniques. Finally, the (15)N-(13)C cross-peak buildup trajectories are purely dipolar in nature (i.e., not influenced by J-couplings or relaxation), which enables the straightforward extraction of (15)N-(13)C(methyl) distances using an analytical model. The SCT-TEDOR experiments are demonstrated on a uniformly (13)C, (15)N-labeled peptide, N-acetyl-Valine, and a 56 amino acid protein, B1 immunoglobulin-binding domain of protein G (GB1), where the measured (15)N-(13)C(methyl) dipolar couplings provide site-specific information about side-chain dihedral angles and the packing of protein molecules in the crystal lattice.

Curr Microbiol. 2008 May; 56(5): 436-41
Singh AK, Syiem MB, Singh RS, Adhikari S, Rai AN

We present evidence, for the first time, of the occurrence of a transport system common for amino acid methionine, and methionine/glutamate analogues L-methionine-DL-sulfoximine (MSX) and phosphinothricin (PPT) in cyanobacterium Nostoc muscorum. Methionine, which is toxic to cyanobacterium, enhanced its nitrogenase activity at lower concentrations. The cyanobacterium showed a biphasic pattern of methionine uptake activity that was competitively inhibited by the amino acids alanine, isoleucine, leucine, phenylalanine, proline, Valine, glutamine, and asparagine. The methionine/glutamate analogue-resistant N. muscorum strains (MSX-R and PPT-R strains) also showed methionine-resistant phenotype accompanied by a drastic decrease in 35S methionine uptake activity. Treatment of protein extracts from these mutant strains with MSX and PPT reduced biosynthetic glutamine synthetase (GS) activity only in vitro and not in vivo. This finding implicated that MSX- and PPT-R phenotypes may have arisen due to a defect in their MSX and PPT transport activity. The simultaneous decrease in methionine uptake activity and in vitro sensitivity toward MSX and PPT of GS protein in MSX- and PPT-R strains indicated that methionine, MSX, and PPT have a common transport system that is shared by other amino acids as well in N. muscorum. Such information can become useful for isolation of methionine-producing cyanobacterial strains.

Neuropsychobiology. 2007; 56(2-3): 152-8
Nordin C, Gupta RC, Sjödin I

Amino acids, such as Valine, isoleucine and leucine compete with tyrosine and tryptophan for transport into the brain and might thus affect the central serotonin and catecholamine patterns. Furthermore, the excitatory amino acids glutamic acid, aspartic acid and glycine are known to act on the N-methyl-D-aspartate receptor, which is part of the reward system. Based on these facts, we have explored the role of cerebrospinal fluid (CSF) amino acids in pathological gambling. Concentrations of amino acids were determined in CSF obtained from one female and 11 pathological male gamblers and 11 healthy male controls. In an ANCOVA with best subset regression, pathological male gamblers had higher CSF levels of the excitatory glutamic and aspartic acids, as well as of phenylalanine, isoleucine, citrulline and glycine. A negative contribution of glycine in interaction with the neuraxis distance might mirror a reduced spinal supply or an altered elimination of glycine in pathological gamblers. A decreasing CSF gradient from the first (0-6 ml) to the third (13-18 ml) CSF fraction was found for glutamic acid, glycine, leucine, isoleucine, lysine, ornithine and glutamine in both pathological gamblers and healthy controls. A decreasing gradient was found, however, for aspartic acid and phenylalanine in pathological male gamblers. The altered pattern of CSF amino acids in pathological gamblers might exert an influence on central monoamines as well as on N-methyl-D-aspartate receptor function.

Plant Physiol Biochem. 2008 Mar; 46(3): 309-24
Duggleby RG, McCourt JA, Guddat LW

Plants and microorganisms synthesize Valine, leucine and isoleucine via a common pathway in which the first reaction is catalysed by acetohydroxyacid synthase (AHAS, EC 2.2.1.6). This enzyme is of substantial importance because it is the target of several herbicides, including all members of the popular sulfonylurea and imidazolinone families. However, the emergence of resistant weeds due to mutations that interfere with the inhibition of AHAS is now a worldwide problem. Here we summarize recent ideas on the way in which these herbicides inhibit the enzyme, based on the 3D structure of Arabidopsis thaliana AHAS. This structure also reveals important clues for understanding how various mutations can lead to herbicide resistance.

J Bacteriol. 2008 Mar; 190(6): 2198-205
Hendrickson EL, Liu Y, Rosas-Sandoval G, Porat I, Söll D, Whitman WB, Leigh JA

Continuous culture, transcriptome arrays, and measurements of cellular amino acid pools and tRNA charging levels were used to determine the response of Methanococcus maripaludis to leucine limitation. For comparison, the responses to phosphate and H2 limitations were measured as well. In addition, the effect of growth rate was determined. leucine limitation resulted in a broad response. tRNA(Leu) charging decreased, but only small increases in mRNA were seen for amino acid biosynthesis genes. However, the cellular levels of free isoleucine and valine showed significant increases, indicating a coordinate regulation of branched-chain amino acids at a post-mRNA level. leucine limitation also resulted in increased mRNA abundance for ribosomal protein genes, increased rRNA abundance, and decreased mRNA abundance for genes of methanogenesis. In contrast, phosphate limitation induced a specific response, a marked increase in mRNA levels for a phosphate transporter. Some mRNA levels responded to more than one factor; for example, transcripts for flagellum synthesis genes decreased under conditions of leucine limitation and increased under H2 limitation. Increased growth rate resulted in increased mRNA levels for ribosomal protein genes, increased rRNA abundance, and increased mRNA for a gene encoding an S-layer protein.