Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.5 (5'-nucleotidase)
 3,167 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A cDNA was cloned coding for human placental 5'-nucleotidase. The 3547-bp cDNA contains an open reading frame that encodes a 574-residue polypeptide with calculated size of 63 375 Da. The NH2-terminal 26 residues comprise a signal peptide, which is followed by the NH2-terminal sequence of the purified protein. four potential N-linked glycosylation sites are found in the molecule, accounting for a larger mass of the mature form (71 kDa). The predicted structure contains a hydrophobic amino acid sequence at the COOH terminus, a possible signal for the post-translational modification by glycophospholipid. To confirm this possibility, we tried to isolate and characterize the membrane-anchoring domain of 5'-nucleotidase. BrCN-cleaved fragments of the protein were extracted with hexane and subjected to HPLC, resulting in purification of a single component of 2.3 kDa. Chemical analyses revealed that the purified fragment contains the tetradecapeptide Lys-Val-Ile-Tyr-Pro-Ala-Val-Glu-Gly-Arg-Ile-Lys-Phe-Ser, ethanolamine, glucosamine, mannose, inositol, palmitic acid, and stearic acid. The peptide sequence determined is identified at positions 510-523 in the primary structure deduced from the cDNA sequence, which predicts a further extension to position 548, containing the hydrophobic amino acid sequence. Thus, it is concluded that the mature 5'-nucleotidase lacks the predicted COOH-terminal peptide extension (524-548), which has been replaced by the glycophospholipid functioning as the membrane anchor of 5'-nucleotidase.
 ...
PMID:Primary structure of human placental 5'-nucleotidase and identification of the glycolipid anchor in the mature form. 212 26

The involvement of glycosylphosphatidylinositol (GPI) in membrane anchoring of 5'-nucleotidase was investigated by chemical analyses. 5'-Nucleotidase purified from rat liver microsomes was subjected to BrCN cleavage, hexane extraction, and high-performance liquid chromatography, resulting in the purification of a single fragment with Mr 2300. Chemical analyses revealed that the purified fragment contains the tetradecapeptide Lys-Val-Ile-Tyr-Pro-Ala-Val-Glu-Gly-Arg-Ile-Lys-Phe-Ser and characteristic components of GPI including ethanolamine, glucosamine, mannose, inositol, palmitic acid, and stearic acid. In addition, it was confirmed that digestion of 5'-nucleotidase with lysyl endopeptidase yielded a fragment containing the dipeptide Phe-Ser and the same GPI components as above. The sequences of the tetradeca- and dipeptides thus determined are identified at positions 510-523 and 522-523, respectively, in the primary structure deduced from the cDNA sequence, which predicts a further extension to position 548, containing a hydrophobic amino acid sequence [Misumi, Y., Ogata, S., Hirose, S., & Ikehara, Y. (1990) J. Biol. Chem. 265, 2178-2183]. Taken together, these results indicate that the mature 5'-nucleotidase molecule lacks the predicted COOH-terminal peptide extension and is attached at serine-523 with GPI, which functions as the membrane anchor of 5'-nucleotidase.
 ...
PMID:Membrane-anchoring domain of rat liver 5'-nucleotidase: identification of the COOH-terminal serine-523 covalently attached with a glycolipid. 214 14

Among the biological exposure indices of lead, lead in plasma was the most direct indicator of current exposure. Lead mobilized into plasma as well as in urine could be used as an indicator of the internal dose of lead. The ratio of non-treated to restored activity of delta-aminolevulinic acid dehydratase (ALA-D) was a more specific index than ALA-D activity itself at low levels of lead exposure, excluding the familial or genetic variation in the activity. The methods using HPLC for determining heme intermediate improved the evaluation of the lead effect: delta-aminolevulinic acid in plasma, blood, and urine (ALA-P, ALA-B, and ALA-U), coproporphyrin in urine, and zinc protoporphyrin in blood (ZP). ROC (Receiver operating characteristic) curve analyses indicated that the diagnostic values for lead exposure decreased in the order ALA-D ratio > ALA-D activity = ALA-P > ALA-U = ZP. Pyrimidine 5'-nucleotidase activity or pyrimidine nucleotide concentrations in blood was also useful for the monitoring or diagnosis of lead intoxication. Using the HPLC method with inclusion compounds in the mobile phase, hippuric acid, methylhippuric acids, mandelic acid and phenylglyoxylic acid could be simultaneously determined in the urine of workers exposed to a mixture of toluene, xylenes, and ethylbenzene. The correction of the urinary metabolite concentration for specific gravity or creatinine allowed the more specific evaluation of the solvent exposure. In the biological monitoring of chlorinated hydrocarbons such as trichloroethylene, prolonged excretion of the metabolites resulted in a bias between metabolite concentrations and TWA levels of the solvent in a day. The background levels of 2,5-hexanedione (HD) were affected by acid hydrolysis conditions, age, sex and lipid metabolism. Substances hydrolyzed to HD in urine from non-exposed subjects were different from HD detected in the workers exposed to n-hexane. Urinary concentrations of N-acetyl-S-(N-methylcarbamoyl) cysteine (AMCC) served as an index of the average exposure to N, N-dimethylformamide during several preceding work days and may indicate the internal dose, while N-methylformamide may be an index of daily exposure. A simple and rapid method for the determination of urinary alkoxyacetic acids was recently developed for the biological monitoring of workers exposed to glycolethers and their acetates. Urinary butoxy acetic acid (free plus conjugated ones) could be simply determined by gaschromatography after acid hydrolysis of urine. The urinary acetone or methanol concentration determined by the head space technique was also useful for the biological monitoring of workers exposed to isopropanol and/or acetone, or methanol, respectively. Evaluation of exposure to the solvents described above could be carried out by comparing the urinary metabolite concentrations with reference values and the biological exposure index values which were defined as the urinary metabolite concentration corresponding to the threshold value for each solvent.
 ...
PMID:[Studies on the evaluation of exposure to industrial chemicals]. 868 99