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1.
Microbiol Spectr ; 10(5): e0189922, 2022 10 26.
Article in English | MEDLINE | ID: mdl-36197290

ABSTRACT

Maintaining the health of seafarers is a difficult task during long-term voyages. Little is known about the corresponding changes in the gut microbiome-host interaction. This study recruited 30 seafarers undertaking a 6-month voyage and analyzed their gut microbiota using 16S rRNA gene sequencing. Fecal untargeted metabolomics analysis was performed using liquid chromatography-mass spectrometry. Significant changes in the composition of the gut microbiota and an increased ratio of Firmicutes/Bacteroidetes at the end (day 180) of the 6-month voyage, relative to the start (day 0), were observed. At the genus level, the abundances of Holdemanella and Plesiomonas were significantly increased, while the abundance of Bacteroides was decreased. Predicted microbial functional analysis revealed significant decreases in folate biosynthesis and biotin metabolism. Furthermore, 20 differential metabolites within six differentially enriched human metabolic pathways (including arginine biosynthesis, lysine degradation, phenylalanine metabolism, sphingolipid metabolism, pentose and glucuronate interconversions, and glycine, serine, and threonine metabolism) were identified by comparing the fecal metabolites at day 0 and day 180. Spearman correlation analysis revealed close relationships between the 14 differential microbiota members and the six differential fecal metabolites that might affect specific human metabolic pathways. This study adopted a multi-omics approach and provides potential targets for maintaining the health of seafarers during long-term voyages. These findings are worthy of more in-depth exploration in future studies. IMPORTANCE Maintaining the health of seafarers undertaking long-term voyages is a difficult task. Apart from the alterations in the gut microbiome and fecal metabolites after a long-term voyage, our study also revealed that 20 differential metabolites within six differentially enriched human metabolic pathways are worthy of attention. Moreover, we found close relationships between the 14 differential microbiota members and the six differential fecal metabolites that might impact specific human metabolic pathways. Accordingly, preventative measures, such as adjusting the gut microbiota by decreasing potential pathobionts or increasing potential probiotics as well as offsetting the decrease in B vitamins and beneficial metabolites (e.g., d-glucuronic acid and citrulline) via dietary adjustment or nutritional supplements, might improve the health of seafarers during long-term sea voyages. These findings provide valuable clues about gut microbiome-host interactions and propose potential targets for maintaining the health of seafarers engaged in long-term sea voyages.


Subject(s)
Gastrointestinal Microbiome , Vitamin B Complex , Humans , Gastrointestinal Microbiome/genetics , RNA, Ribosomal, 16S/genetics , Vitamin B Complex/analysis , Citrulline/analysis , Biotin , Lysine/analysis , Metabolomics/methods , Feces , Pentoses/analysis , Glucuronates/analysis , Glycine/analysis , Glucuronic Acid , Serine/analysis , Phenylalanine/analysis , Sphingolipids/analysis , Threonine/analysis , Arginine/analysis , Folic Acid/analysis
2.
Metabolism ; 102: 154000, 2020 01.
Article in English | MEDLINE | ID: mdl-31678070

ABSTRACT

OBJECTIVE: Supplementation with serine attenuates alcoholic fatty liver by regulating homocysteine metabolism and lipogenesis. However, little is known about serine metabolism in fatty liver disease (FLD). We aimed to investigate the changes in serine biosynthetic pathways in humans and animal models of fatty liver and their contribution to the development of FLD. METHODS: High-fat diet (HFD)-induced steatosis and methionine-choline-deficient diet-induced steatohepatitis animal models were employed. Human serum samples were obtained from patients with FLD whose proton density fat fraction was estimated by magnetic resonance imaging. 3-Phosphoglycerate dehydrogenase (Phgdh)-knockout mouse embryonic fibroblasts (MEF) and transgenic mice overexpressing Phgdh (Tg-phgdh) were used to evaluate the role of serine metabolism in the development of FLD. RESULTS: Expression of Phgdh was markedly reduced in the animal models. There were significant negative correlations of the serum serine with the liver fat fraction, serum alanine transaminase, and triglyceride levels among patients with FLD. Increased lipid accumulation and reduced NAD+ and SIRT1 activity were observed in Phgdh-knockout MEF and primary hepatocytes incubated with free fatty acids; these effects were reversed by overexpression of Phgdh. Tg-Phgdh mice showed significantly reduced hepatic triglyceride accumulation compared with wild-type littermates fed a HFD, which was accompanied by increased SIRT1 activity and reduced expression of lipogenic genes and proteins. CONCLUSIONS: Human and experimental data suggest that reduced Phgdh expression and serine levels are closely associated with the development of FLD.


Subject(s)
Glyceraldehyde-3-Phosphate Dehydrogenases/genetics , Liver/metabolism , Non-alcoholic Fatty Liver Disease/genetics , Non-alcoholic Fatty Liver Disease/metabolism , Serine/metabolism , Animals , Cells, Cultured , Cohort Studies , Diet, High-Fat , Down-Regulation , Embryo, Mammalian , Female , Gene Expression Regulation, Enzymologic , Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism , Humans , Lipid Metabolism/genetics , Lipogenesis/genetics , Liver/chemistry , Liver/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Non-alcoholic Fatty Liver Disease/pathology , Serine/analysis
3.
J Dairy Sci ; 102(8): 6781-6789, 2019 Aug.
Article in English | MEDLINE | ID: mdl-31155253

ABSTRACT

Selenium is included in selenoprotein sequences, which participate in enzymatic processes necessary to preserve optimal health. Some lactic acid bacteria carry out the biotransformation of inorganic selenium in their metabolism. The complete biochemical mechanism of selenium biotransformation is still unknown; however, it is known that both the selenocysteine synthesis process and its subsequent incorporation into selenoproteins include serine as part of the action of seryl-RNAt synthetase. Therefore, the aim of this work was to determine the effect of serine during the biotransformation of selenium and the subsequence growth of Streptococcus thermophilus in a minimal medium. Two culture media were prepared, one enriched with the minimum inhibitory concentration of selenite (as Na2SeO3) and the other as a mixture of the minimum inhibitory concentration of selenite and serine. The absorbed selenium concentration was measured by inductively coupled plasma, and the selenocysteine identification was performed by reverse-phase HPLC. In the second culture medium, decreases in both times, the adaptation and the logarithmic phase, were observed. According to the results, it was possible to establish that the presence of serine allowed the biotransformation of selenite into selenocysteine by Strep. thermophilus.


Subject(s)
Culture Media/chemistry , Selenium/metabolism , Selenocysteine/biosynthesis , Serine/administration & dosage , Streptococcus thermophilus/metabolism , Animals , Chromatography, High Pressure Liquid , Selenoproteins , Serine/analysis
4.
Anal Chem ; 90(10): 6043-6050, 2018 05 15.
Article in English | MEDLINE | ID: mdl-29565564

ABSTRACT

Cross-linking/Mass spectrometry (XLMS) is a consolidated technique for structural characterization of proteins and protein complexes. Despite its success, the cross-linking chemistry currently used is mostly based on N-hydroxysuccinimide (NHS) esters, which react primarily with lysine residues. One way to expand the current applicability of XLMS into several new areas is to increase the number of cross-links obtainable for a target protein. We introduce a multiplex chemistry (denoted XPlex) that targets Asp, Glu, Lys, and Ser residues. XPlex can generate significantly more cross-links with reactions occurring at lower temperatures and enables targeting proteins that are not possible with NHS ester-based cross-linkers. We demonstrate the effectiveness of our approach in model proteins as well as a target Lys-poor protein, SalBIII. Identification of XPlex spectra requires a search engine capable of simultaneously considering multiple cross-linkers on the same run; to achieve this, we updated the SIM-XL search algorithm with a search mode tailored toward XPlex. In summary, we present a complete chemistry/computational solution for significantly increasing the number of possible distance constraints by mass spectrometry experiments, and thus, we are convinced that XPlex poses as a real complementary approach for structural proteomics studies.


Subject(s)
Aspartic Acid/analysis , Computational Biology , Cross-Linking Reagents/chemistry , Glutamic Acid/analysis , Lysine/analysis , Serine/analysis , Algorithms , Esters/chemistry , Mass Spectrometry , Proteins/chemistry , Succinimides/chemistry , Temperature
5.
Zhong Yao Cai ; 35(5): 686-9, 2012 May.
Article in Chinese | MEDLINE | ID: mdl-23213726

ABSTRACT

OBJECTIVE: To improve the quality standard of Scolopendra subspinipes mutilans by researching the methods of the TLC identification and anti-coagulant activity quantitatively. METHODS: Identified the free arginine (Arg) and serine (Ser) in scolopendra by TLC, screened the samples preparation process and developed solvent systems; Determined the anti-coagulant activity by method of titration with thrombin and screened the pretreatment methods. RESULTS: When medicinal materials was extracted by formic acid and 95% ethanol (1:1) with ultrasonic method and developed by n-butanol-acetic acid-water (12:5:4), the spots of Arg and Ser were well separated. Ultrasonic method was suitable for preparation of the anti-coagulant components in Scolopendra subspinipes mutilans and their anti-coagulant activity was determined by method of titration with thrombin could get a well reproducibility, the anti-thrombin activity of testing sample was (14.00 +/- 1.53) U/g and those of three different batch were (13.00 +/- 0.58) U/g, (17.00 +/- 1.15) U/g, (15.67 +/- 1.53) U/g respectively. CONCLUSION: The methods of TLC identification and anti-coagulant activity quantitatively could be used as a basis for improving the quality standard of Scolopendra subspinipes mutilans.


Subject(s)
Anticoagulants/pharmacology , Arthropods , Materia Medica/chemistry , Materia Medica/pharmacology , Thrombin/analysis , Animals , Anticoagulants/chemistry , Anticoagulants/isolation & purification , Arginine/analysis , Arthropods/chemistry , Chromatography, Thin Layer/methods , Materia Medica/isolation & purification , Quality Control , Reproducibility of Results , Serine/analysis , Technology, Pharmaceutical/methods , Thrombin/antagonists & inhibitors , Titrimetry/methods
6.
Dev Dyn ; 236(2): 374-88, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17191254

ABSTRACT

The beta-keratins constitute the hard epidermis and adhesive setae of gecko lizards. Nucleotide and amino acid sequences of beta-keratins in epidermis of gecko lizards were cloned from mRNAs. Specific oligonucleotides were used to amplify by 3'- and 5'-rapid amplification of cDNA ends analyses five specific gecko beta-keratin cDNA sequences. The cDNA coding sequences encoded putative glycine-proline-serine-rich proteins of 16.8-18 kDa containing 169-191 amino acids, especially 17.8-23% glycine, 8.4-14.8% proline, 14.2-18.1% serine. Glycine-rich repeats are localized toward the initial and end regions of the protein, while a central region, rich in proline, has a strand conformation (beta-pleated fold) likely responsible for the formation of beta-keratin filaments. It shows high homology with a core region of other lizard keratins, avian scale, and feather keratins. Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis show a higher beta-keratin gene expression in regenerating epidermis compared with normal epidermis. In situ hybridization confirms that mRNAs for these proteins are expressed in cells of the differentiating oberhautchen cells and beta-cells. Expression in adhesive setae of climbing lamellae was shown by RT-PCR. Southern blotting analysis revealed that the proteins are encoded by a multigene family. PCR analysis showed that the genes are presumably located in tandem along the DNA and are transcribed from the same DNA strand like in avian beta-keratins.


Subject(s)
Amino Acid Motifs/genetics , Epidermis/chemistry , Gene Expression , Lizards/embryology , Lizards/genetics , beta-Keratins/genetics , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Blotting, Southern , Cloning, Molecular , DNA, Complementary/genetics , Glycine/analysis , Immunohistochemistry , In Situ Hybridization , Molecular Sequence Data , Oligonucleotides , Proline/analysis , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology , Serine/analysis , beta-Keratins/chemistry , beta-Keratins/metabolism
7.
J Neurophysiol ; 96(1): 465-70, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16687621

ABSTRACT

Direct interactions between the presynaptic N-type calcium channel and the beta subunit of the heterotrimeric G-protein complex cause voltage-dependent inhibition of N-type channel activity, crucially influencing neurotransmitter release and contributing to analgesia caused by opioid drugs. Previous work using chimeras of the G-protein beta subtypes Gbeta1 and Gbeta5 identified two 20-amino acid stretches of structurally contiguous residues on the Gbeta1 subunit as critical for inhibition of the N-type channel. To identify key modulation determinants within these two structural regions, we performed scanning mutagenesis in which individual residues of the Gbeta1 subunit were replaced by corresponding Gbeta5 residues. Our results show that Gbeta1 residue Ser189 is critical for N-type calcium channel modulation, whereas none of the other Gbeta1 mutations caused statistically significant effects on the ability of Gbeta1 to inhibit N-type channels. Structural modeling shows residue 189 is surface exposed, consistent with the idea that it may form a direct contact with the N-type calcium channel alpha1 subunit during binding interactions.


Subject(s)
Calcium Channels, N-Type/physiology , GTP-Binding Protein beta Subunits/chemistry , GTP-Binding Protein beta Subunits/physiology , Serine/analysis , Serine/physiology , Cell Line , DNA, Complementary/analysis , DNA, Complementary/genetics , Electrophysiology , G Protein-Coupled Inwardly-Rectifying Potassium Channels/physiology , GTP-Binding Protein beta Subunits/genetics , Humans , Mutagenesis , Patch-Clamp Techniques , Protein Structure, Tertiary
8.
Poult Sci ; 83(8): 1314-24, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15339006

ABSTRACT

Male chickens of a broiler (B) and a layer (L) genotype were grown in floor pens from d 8 to 21 posthatch in groups of 10. Three pens per genotype were allocated to each of 10 experimental diets. The diets were offered ad libitum and they differed in lysine concentration from 3.8 to 16.8 g/kg. The source of supplemental lysine was L-lysine x HCl. All birds were killed at the end of the experiment, and representative birds (3 groups of 10 per genotype) were killed at the start for baseline measurements. Accretions of protein, fat, energy, and amino acids were determined by comparative body analysis. Responses were described with sigmoidal and exponential functions. Additionally, the net disappearance rate (NDR) of amino acids from the small intestine was studied with the basal diet (3.8 g of lysine/kg) using 6 replicated pens of 15 birds per genotype. Titanium dioxide was the indigestible marker. Net disappearance rates were not significantly different between genotypes for CP or any amino acid. Responses to incremental lysine concentration were nonlinear for both genotypes but distinctly different in magnitude between genotypes. Estimated y(max) values for 14-d BW, protein gain, and gain/ feed ratio were 534 (B) and 153 (L) g, 87.1 (B) and 28.7 (L) g, and 0.82 (B) and 0.71 (L) g/g. Protein accretion approached 95% of the estimated y(max) with dietary lysine concentrations of 12.5 (B) and 10.4 (L) g/kg. The amino acid profile of accreted whole body protein was different between genotypes, and was affected by supplementary lysine. Lysine content in accreted whole body protein approached upper values of 7.4 (B) and 5.6 (L) g/16 g of N with increasing dietary lysine concentration. Marginal efficiency of lysine utilization, determined as delta lysine accretion/delta lysine intake, showed maxima of 99% (B) and 74% (L). These maxima were achieved at intakes which were much lower than those needed for high protein accretion. It was concluded that the efficiency of amino acid utilization may depend on genotype, perhaps due to differences in the relative proportion of different protein fractions to whole body protein and due to differences in the ratio of synthesis and degradation of body proteins. Nonlinear relationships and different amino acid pattern of accreted body protein should be implemented in future models of requirements.


Subject(s)
Body Composition , Chickens/growth & development , Chickens/genetics , Lysine/administration & dosage , Lysine/metabolism , Weight Gain , Amino Acids/metabolism , Animals , Cysteine/analysis , Energy Metabolism , Genotype , Glycine/analysis , Isoleucine/analysis , Lipid Metabolism , Male , Methionine/analysis , Proline/analysis , Proteins/analysis , Proteins/metabolism , Serine/analysis
9.
Bull Exp Biol Med ; 137(2): 167-70, 2004 Feb.
Article in English, Russian | MEDLINE | ID: mdl-15273766

ABSTRACT

Amino acid composition of a new Russian nephroprotective drug Nephrophyt (a complex mixture of dry plant extracts) was studied for the first time in order to validate its pharmacological effects in rats with mercuric chloride-induced nephropathy. The concentration of pyrrolidone amino acids and serine in this preparation was extremely high, which is uncommon for plant raw material. These data are in good correlation with pronounced nephroprotective effect and agree with published reports on pharmacological effects of individual amino acids and their metabolites. The prospects of the use of this preparation based on the data on its amino acid composition are discussed.


Subject(s)
Acute Kidney Injury/drug therapy , Phytotherapy , Plant Preparations/chemistry , Plant Preparations/pharmacology , Acute Kidney Injury/chemically induced , Amino Acids/analysis , Animals , Female , Male , Mercuric Chloride/toxicity , Rats , Serine/analysis
10.
FEBS Lett ; 528(1-3): 125-9, 2002 Sep 25.
Article in English | MEDLINE | ID: mdl-12297291

ABSTRACT

A peptide named apisimin was found in honeybee (Apis mellifera L.) royal jelly (RJ). N-terminal sequencing showed that this peptide corresponded to the sequence of a cDNA clone isolated from an expression cDNA library prepared from heads of nurse honeybees. No homology was found between the protein sequence of apisimin with a molecular mass of 5540.4 Da and sequences deposited in the Swiss-Prot database. The 54 amino acids of apisimin do not include Cys, Met, Pro, Arg, His, Tyr, and Trp residues. The peptide shows a well-defined secondary structure as observed by CD spectroscopy, and has the tendency to form oligomers. Isoelectrofocusing showed apisimin to be an acidic peptide.


Subject(s)
Bees/chemistry , Fatty Acids/chemistry , Insect Proteins/isolation & purification , Amino Acid Sequence , Animals , Base Sequence , Bees/genetics , Circular Dichroism , DNA, Complementary/genetics , Insect Proteins/chemistry , Insect Proteins/genetics , Molecular Sequence Data , Molecular Weight , Protein Structure, Secondary , Serine/analysis , Valine/analysis
11.
Biochim Biophys Acta ; 1447(1): 77-92, 1999 Oct 06.
Article in English | MEDLINE | ID: mdl-10500247

ABSTRACT

In order to sequence the cysteine-rich regions of pig gastric mucin (PGM), we used our previously identified pig gastric mucin clone PGM-2A to screen a pig stomach cDNA library and perform rapid amplification of cDNA ends to obtain two cysteine-rich clones, PGM-2X and PGM-Z13. PGM-2X has 1071 base pairs (bp) encoding 357 amino acids containing five serine-threonine-rich 16 amino acid tandem repeats, downstream from a cysteine-rich region similar to human and mouse MUC5AC. PGM-Z13 encodes the complete 3'-terminus of PGM and is composed of 3336 bp with a 2964 bp open reading frame encoding 988 amino acids with four serine-threonine-rich tandem repeats upstream from a cysteine-rich region similar to the carboxyl terminal regions of human and rat MUC5AC and human MUC5B. This region is homologous to von Willebrand factor C and D domains involved in acid induced polymerization, and to the carboxyl terminal cystine-knot domain of various mucins, TGF-beta, vWF and norrin, which is involved in dimerization. These newly sequenced cysteine-rich regions of pig gastric mucin may be critical for its gelation and for its observed increased viscosity induced by low pH.


Subject(s)
Cysteine/analysis , Cystine/analysis , Gastric Mucins/chemistry , von Willebrand Factor/chemistry , Amino Acid Sequence , Animals , Base Sequence , Cystine/genetics , DNA, Complementary/chemistry , Epithelium/chemistry , Gastric Mucosa/chemistry , Hydrogen-Ion Concentration , Molecular Sequence Data , Sequence Alignment , Serine/analysis , Swine , Tandem Repeat Sequences , Threonine/analysis , von Willebrand Factor/genetics
12.
Plant Cell ; 10(10): 1637-48, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9761791

ABSTRACT

The U1 small nuclear ribonucleoprotein particle (U1 snRNP) 70K protein (U1-70K), one of the three U1 snRNP-specific proteins, is implicated in basic and alternative splicing of nuclear pre-mRNAs. We have used the Arabidopsis U1-70K in the yeast two-hybrid system to isolate cDNAs encoding proteins that interact with it. This screening has resulted in the isolation of two novel plant serine/arginine-rich (SR) proteins, SRZ-22 and SRZ-21 (SRZ proteins). Neither the N-terminal region nor the arginine-rich C-terminal region of U1-70K alone interact with the SRZ proteins. The interaction of U1-70K with the SRZ proteins is confirmed further in vitro using a blot overlay assay. The plant SRZ proteins are highly similar to each other and contain conserved modular domains unique to different groups of splicing factors in the SR family of proteins. SRZ proteins are similar to human 9G8 splicing factor because they contain a zinc knuckle, precipitate with 65% ammonium sulfate, and cross-react with the 9G8 monoclonal antibody. However, unlike the 9G8 splicing factor, SRZ proteins contain a glycine hinge, a unique feature in other splicing factors (SC35 and ASF/SF2), located between the RNA binding domain and the zinc knuckle. SRZ-22 and SRZ-21 are encoded by two distinct genes and are expressed in all tissues tested with varied levels of expression. Our results suggest that the plant SRZ proteins represent a new group of SR proteins. The interaction of plant U1-70K with the SRZ proteins may account for some differences in pre-mRNA splicing between plants and animals.


Subject(s)
Plant Proteins/metabolism , Ribonucleoprotein, U1 Small Nuclear/metabolism , Amino Acid Sequence , Animals , Arabidopsis/genetics , Arabidopsis/metabolism , Arginine/analysis , Base Sequence , DNA, Complementary/genetics , DNA, Plant/genetics , Escherichia coli/genetics , Gene Expression , Genes, Plant , Humans , Molecular Sequence Data , Plant Proteins/chemistry , Plant Proteins/genetics , RNA Precursors/genetics , RNA Precursors/metabolism , RNA Splicing , RNA, Plant/genetics , RNA, Plant/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Saccharomyces cerevisiae/genetics , Sequence Homology, Amino Acid , Serine/analysis
13.
Connect Tissue Res ; 33(1-3): 67-72, 1995.
Article in English | MEDLINE | ID: mdl-7554964

ABSTRACT

Acidic phosphorylated proteins are prominent constituents of the extracellular matrix of bone and dentin. It has been postulated that they may have important structural and regulatory roles in the process of tissue mineralization. Studies of a cDNA library, prepared from cells of the rat incisor odontoblast-pulp complex of 3 week old Sprague-Dawley rats, led to the identification of a serine-rich acidic protein, designated AG1, which appeared to be a dentin matrix component. In order to determine which cells of the odontoblast-pulp complex were responsible for the making of AG1, in situ hybridization was carried out using digoxigenin-labeled probes. The full length AG1 cDNA was subcloned into the pBluescript vector, which contains two strong promoters, T3 and T7. The sense and antisense complementary RNA (cRNA) hybridization probes were prepared by in vitro transcription using T3 and T7 polymerases in the presence of 11-dUTP. Incisor sections were obtained from rat embryos at days 16, and 20, and newborns at days 2 and 5. No AG1 mRNA was detected in the embryonic sections, but digoxigenin labeling was evident in odontoblasts secreting mineralizing dentin at postnatal days 2 and 5. Sense probes showed no hybridization. Pulp cells, Meckel's cartilage, and alveolar bone were free of hybridization with the antisense probe. Unexpectedly, a low level of digoxigenin staining was seen in the cytoplasm of secretory ameloblasts, but not in the preameloblasts, stratum intermedium or stellate reticulum of the enamel organ. These data show that AG1 expression is regulated developmentally and is restricted to secretory stage mature odontoblasts.


Subject(s)
Dentin/metabolism , Extracellular Matrix Proteins/analysis , Odontoblasts/metabolism , Phosphoproteins/analysis , Alveolar Process/cytology , Alveolar Process/metabolism , Ameloblasts/metabolism , Animals , Cartilage/cytology , Cartilage/metabolism , DNA Probes , DNA, Complementary/genetics , Dental Pulp/cytology , Dental Pulp/metabolism , Dentin/cytology , Digoxigenin , Extracellular Matrix Proteins/genetics , Gene Expression Regulation, Developmental , Genetic Vectors , In Situ Hybridization , Phosphoproteins/genetics , Promoter Regions, Genetic/genetics , RNA Probes , RNA, Antisense/genetics , RNA, Complementary/genetics , Rats , Rats, Sprague-Dawley , Serine/analysis , Serine/genetics , Tooth Calcification , Transcription, Genetic
14.
Neurosci Lett ; 120(2): 237-40, 1990 Dec 11.
Article in English | MEDLINE | ID: mdl-2127307

ABSTRACT

We studied the kinetics of the enzyme serine hydroxymethyl transferase (SHMT) and the concentration of its metabolic substrates serine and glycine, in the postmortem brains of controls and schizophrenics. The Km of SHMT, and the concentration of serine and glycine were all significantly higher in the temporal lobes of brain tissues from schizophrenics than in those from controls. These differences were not observed in the frontal lobe specimens. Neuroleptics, age, sex and autolysis time did not contribute to these differences. The role of SHMT deficiency in schizophrenia is discussed in relation to the production of glycine and 1-carbon units from which purines and thereby adenosine is produced. Both glycine and adenosine are potent neuromodulatory substances for the release of dopamine and glutamate, neurotransmitters which have been implicated in the pathophysiology of schizophrenia.


Subject(s)
Glycine Hydroxymethyltransferase/metabolism , Schizophrenia/enzymology , Temporal Lobe/enzymology , Adult , Female , Frontal Lobe/chemistry , Frontal Lobe/metabolism , Glycine/analysis , Humans , Male , Middle Aged , Reference Values , Serine/analysis , Temporal Lobe/chemistry , gamma-Aminobutyric Acid/analysis
15.
Neuropharmacology ; 25(7): 703-9, 1986 Jul.
Article in English | MEDLINE | ID: mdl-3092127

ABSTRACT

Concentrations of gamma-aminobutyric acid (GABA), glycine and serine have been measured in 44 microdissected areas of the brain of the rat. All three amino acids were ubiquitously present and distributed unevenly in the brain. Very high levels of GABA were found in the anterior hypothalamic and medial preoptic nuclei and the substantia nigra; high levels were found in the interpeduncular and red nuclei in the mesencephalon and in several hypothalamic nuclei. Glycine was distributed fairly uniformly with large concentrations in certain lower brainstem nuclei. In these areas, the concentrations of glycine exceeded those of serine, while the serine-glycine ratio was 4.5:1 in the caudate nucleus, 4:1 in the cerebellum and 2.5:1 in the cerebral cortical areas. Acute stress induced with formalin (pain) resulted in a significant depletion of levels of GABA in the hypothalamus and the lower brainstem but not in the cortical areas. In the same animals, concentrations of glycine doubled in the cerebral cortex and remained unchanged elsewhere in the brain. Increased motor and behavioral activity after the acute administration of a large dose of amphetamine were associated with a 2-5-fold increase in the levels of glycine in brain, and markedly elevated the concentrations of GABA in the major biogenic amine-containing cell groups only (substantia nigra, locus coeruleus and dorsal raphe).


Subject(s)
Brain Chemistry , Glycine/analysis , gamma-Aminobutyric Acid/analysis , Animals , Male , Postmortem Changes , Rats , Rats, Inbred Strains , Serine/analysis
16.
J Neurochem ; 45(6): 1802-8, 1985 Dec.
Article in English | MEDLINE | ID: mdl-2865334

ABSTRACT

Sprague-Dawley dams were fed either a protein-calorie deficient or control diet from day 5 to day 21 after parturition. The concentrations of seven amino acids (aspartate, glutamate, gamma-aminobutyric acid, glycine, glutamine, serine, and taurine) were determined in brain regions from 17-day-old undernourished offspring and from 35-day-old rehabilitated rats. The brain regions examined were the cortex, cerebellum, corpus striatum, hippocampus, hypothalamus, brainstem, and midbrain. At 17 days of age, taurine was the amino acid with the highest concentration, whereas at 35 days glutamate had the highest concentration. This change was due to the fact that the concentration of taurine decreased significantly in all brain regions between 17 and 35 days, whereas the concentration of glutamate remained high or increased somewhat in all brain regions except the hypothalamus and brainstem. When the age-matched offspring of control and undernourished rats were compared, several interesting and significant differences were found. The concentrations of glutamate and aspartate were significantly lower (decreased 16-34%) in the cerebellum, brainstem, cortex, and midbrain in 17-day-old undernourished rats. The aspartate level was also significantly decreased in the corpus striatum and hypothalamus in 17-day-old offspring. However, the deficiencies of aspartate and glutamate were transient and reversible. In contrast, the concentration of taurine was increased in the hypothalamus (31%) and hippocampus (12-33%) at both 17 and 35 days of age and in the midbrain (17%) at 17 days. Other transient abnormalities in amino acid levels were found in undernourished offspring. The results of these experiments suggest that undernutrition during lactation causes delayed CNS development, which is manifested in altered concentrations of the neurotransmitters aspartate, glutamate, and taurine.


Subject(s)
Amino Acids/analysis , Brain Chemistry , Lactation , Nutrition Disorders/metabolism , Animals , Aspartic Acid/analysis , Body Weight , Corpus Striatum/analysis , Female , Glutamates/analysis , Glutamic Acid , Glutamine/analysis , Glycine/analysis , Hippocampus/analysis , Hypothalamus/analysis , Pregnancy , Rats , Rats, Inbred Strains , Serine/analysis , Taurine/analysis , gamma-Aminobutyric Acid/analysis
20.
J Virol ; 23(1): 196-204, 1977 Jul.
Article in English | MEDLINE | ID: mdl-69721

ABSTRACT

The pP60gag polyprotein of the feline leukemia virus pseudotype of m1 Moloney murine sarcoma virus [m1MSV(FeLV)] was previously shown to be MSV specific and to contain murine p30 and smaller structural polypeptides. This protein was detected in m1MSV-transformed cells, and in pulse-chase studies it was found to be stable. In this study virion P60 was shown to contain murine pp12, to be phosphorylated, and to bind to nucleic acids. 32P-labeled m1MSV[FeLV) was fractionated by guanidine agarose chromatography and analyzed by gel electrophoresis. Both P60 and pp12 were found to be the major phosphoproteins, phosphorylated in both serine and threonine residues. Virion P60 bound preferentially to single-stranded DNA and RNA in a competition filter binding assay, using 125I-labeled single-stranded calf thymus DNA and various unlabeled nucleic acids. Similar phosphorylation and DNA binding properties were demonstrated for cellular P60. Thus, immunoprecipitation of cellular extracts showed that P60 was phosphorylated in both producer and nonproducer transformed cells, indicating that phosphorylation occurs independently of virus assembly. Moreover, P60 from cytoplasmic extracts was retained on single-stranded DNA-Sepharose columns, demonstrating that cellular P60 binds to DNA.


Subject(s)
DNA/metabolism , Gammaretrovirus/metabolism , Phosphorus/metabolism , RNA/metabolism , Sarcoma Viruses, Murine/metabolism , Viral Proteins/metabolism , DNA, Single-Stranded/metabolism , Epitopes , Leukemia Virus, Feline , Phosphoproteins/analysis , Phosphoproteins/immunology , Protein Binding , Sarcoma Viruses, Murine/analysis , Sarcoma Viruses, Murine/immunology , Serine/analysis , Threonine/analysis , Viral Proteins/analysis , Viral Proteins/immunology
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