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

Sarcolemmal vesicles isolated from human skeletal muscle obtained at surgery showed approximately 14-fold enrichment of sarcolemmal marker enzymes 5'-nucleotidase and K-stimulated phosphatase. [3H]glutamine transport in these vesicles was stereospecific, largely Na dependent, and tolerated Li-for-Na substitution. Glutamine transport was stimulated by an inside negative membrane potential, and 25 mM glutamine stimulated 22Na (0.1 mM) uptake into vesicles by 50%, indicating rheogenic cotransport of Na and glutamine. Alanine transport was Na dependent but did not tolerate Li-for-Na substitution. Transport of L-[3H]glutamine was inhibited by 35-65% with a 20-fold excess of glutamine, asparagine, and alanine; cysteine, alpha-(methylamino)isobutyrate, and 2-amino-2-norborane carboxylic acid had smaller inhibitory effects, although cysteine had an unusually large inhibitory effect on glutamine transport at 1,000-fold excess compared with most other amino acids. Glutamine transport showed sensitivity to pH values < 7.0. Glutamine transport consisted of a Na-dependent and a Na-independent component, both of which appeared saturable. The kinetic characteristics of the Na-dependent component were different in different types of muscles, with half-maximal concentrations (mM) varying from 1.6 +/- 0.4 (tibialis anterior) to 0.56 +/- 0.0.2 (gluteus maximus) and maximal velocity (pmol.mg protein-1.s-1) of 1.3 +/- 0.27 to 5 +/- 1.25 in the same muscles. The results demonstrate both marked similarities and important differences between the principal glutamine transporter in human skeletal muscle and the known system Nm transporter in rat skeletal muscle.
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PMID:Glutamine transport in human skeletal muscle. 833 25

HD-domain phosphohydrolases have nucleotidase and phosphodiesterase activities and play important roles in the metabolism of nucleotides and in signaling. We present three 2.1-A-resolution crystal structures (one in the free state and two complexed with natural substrates) of an HD-domain phosphohydrolase, the Escherichia coli 5'-nucleotidase YfbR. The free-state structure of YfbR contains a large cavity accommodating the metal-coordinating HD motif (H33, H68, D69, and D137) and other conserved residues (R18, E72, and D77). Alanine scanning mutagenesis confirms that these residues are important for activity. Two structures of the catalytically inactive mutant E72A complexed with Co(2+) and either thymidine-5'-monophosphate or 2'-deoxyriboadenosine-5'-monophosphate disclose the novel binding mode of deoxyribonucleotides in the active site. Residue R18 stabilizes the phosphate on the Co(2+), and residue D77 forms a strong hydrogen bond critical for binding the ribose. The indole side chain of W19 is located close to the 2'-carbon atom of the deoxyribose moiety and is proposed to act as the selectivity switch for deoxyribonucleotide, which is supported by comparison to YfdR, another 5'-nucleotidase in E. coli. The nucleotide bases of both deoxyriboadenosine-5'-monophosphate and thymidine-5'-monophosphate make no specific hydrogen bonds with the protein, explaining the lack of nucleotide base selectivity. The YfbR E72A substrate complex structures also suggest a plausible single-step nucleophilic substitution mechanism. This is the first proposed molecular mechanism for an HD-domain phosphohydrolase based directly on substrate-bound crystal structures.
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PMID:Structural insight into the mechanism of substrate specificity and catalytic activity of an HD-domain phosphohydrolase: the 5'-deoxyribonucleotidase YfbR from Escherichia coli. 1835 68