The dipeptide N-acetylaspartyl-glutamate (NAAG) is an abundant neuropeptide in the mammalian brain. Despite this fact, its physiological role is poorly understood. NAAG is synthesized by a NAAG synthetase catalyzing the ATP-dependent condensation of N-acetylaspartate and glutamate. In vitro NAAG synthetase activity has not been described, and the enzyme has not been purified. Using a bioinforma...

BACKGROUND Phospholemman (PLM) is an important phosphorylation substrate for protein kinases A and C in the heart. Until now, the association between PLM phosphorylation status and L-type calcium channels (LTCCs) gating has not been fully understood. We investigated the kinetics of LTCCs in HEK 293T cells expressing phosphomimetic or nonphosphorylatable PLM mutants. METHODS The LTCCs gating w...

Stop codon readthrough is used extensively by viruses to expand their gene expression. Until recent discoveries in Drosophila, only a very limited number of readthrough cases in chromosomal genes had been reported. Analysis of conserved protein coding signatures that extend beyond annotated stop codons identified potential stop codon readthrough of four mammalian genes. Here we use a modified t...

RNA interference (RNAi) mediated by expression of short hairpin RNAs (shRNAs) is a powerful tool for efficiently suppressing target genes. The approach allows studies of the function of individual genes and may also be applied to human therapy. However, in many instances regulation of RNAi by administration of a small inducer molecule will be required. To date, the development of appropriate re...

Natural resistance-associated macrophage protein 2 (Nramp2) has been suggested to be involved in transferrin-independent iron uptake. Two isoforms of the Nramp2 gene generated by alternative splicing of the 3' exons were identified in mouse, rat and human, but it is unclear if they perform distinct functions. To rationalize our previous work, which indicated an increase in iron deposition in a ...

OBJECTIVE miR-483-5p has been identified as a miRNA oncogene in certain cancers. However, its role in prostate cancer has not been sufficiently investigated. In this study, we investigated the role of miR-483-5p in prostate cancer and examined RBM5 regulation by miR-483-5p. MATERIAL AND METHODS Expression levels of miR-483-5p were determined by quantitative real-time PCR. The effect of miR-48...

Oxovanadium(IV) complexes [VO(aip)(L)](ClO4)2 (L = phtpy, 1; stpy, 2) and [VO(pyip)(L)](ClO4)2 (L = phtpy, 3; stpy, 4), where aip is 2-(9-anthryl)-1H-imidazo[4,5-f][1,10]phenanthroline, pyip is [2-(1-pyrenyl)-1H-imidazo[4,5-f][1,10]phenanthroline, phtpy is (4'-phenyl)-2,2':6',2''-terpyridine and stpy is (2,2':6',2''-terpyridin-4'-oxy)ethyl-β-D-glucopyranoside, were prepared, characterized and t...

Selenoproteins exhibit diverse biological functions, most of which are associated with redox control. However, the functions of approximately half of mammalian selenoproteins are not known. One such protein is Selenoprotein O (SelO), the largest mammalian selenoprotein with orthologs found in a wide range of organisms, including bacteria and yeast. Here, we report characterization of mammalian ...

Reduced glutathione (GSH) is an intracellular molecule essential for many aspects of cell physiology and defense. Determination of GSH has been used to identify potential anti-cancer drugs and for the assessment of drug toxicity via generation of oxidative stress. The described protocol was designed to modify existing protocols for the fluorescent detection of intracellular GSH in a high throug...

Previously we identified MIR16 (membrane interacting protein of RGS16) as an integral membrane glycoprotein that interacts with regulator of G protein signaling proteins and shares significant sequence homology with bacterial glycerophosphodiester phosphodiesterases (GDEs), suggesting that it is a putative mammalian GDE. Here we show that MIR16 belongs to a large, evolutionarily conserved famil...