The Functional assessment of fluorescently tagged adenosine A2A and dopamine D2 receptors and qualitative analysis of dimerization of adenosine A2A and dopamine D2 receptor by using FRET

Download
2013
Akkuzu, Selin
Recently, several studies have demonstrated that G protein coupled receptors exist as homo/heterodimers or oligomers. Adenosine A2A receptors and dopamine D2 receptors are present as both homo- and heterodimer. In the GABAergic striatopallidal neurons A2AR are co- localized with D2 receptors (D2R), and establish functional A2AR-D2R heteromers, which modulates dopaminergic activity. Due to be involved in physiological processes, these receptors bear critical roles. Dopamine receptors play critical role in dopaminergic pathways in regulation of memory, food intake and psychomotor activity, etc. On the other hand, adenosine A2A receptors are involved in the regulations of neurotransmission, immune response and cardiovascular systems. Dopamine D2R andadenosine A2AR have been shown to interact in striatum and modulate dopaminergic activity The purpose of this study is to assess the functionality of EGFP (enhanced green fluorescent protein) and mCherry (a red fluorescent protein) tagged adenosine A2A and dopamine D2 receptors and to detect homo/ hetero-dimerization of these receptors in live cells via Fluorescence Resonance Energy Transfer (FRET). Understanding the mechanisms of the interaction between adenosine and dopamine signaling will help us to figure out some molecular mechanism of neurophysiological disorders. Furthermore, the fluorescence based live cell model could be used to observe the effects of potential anti-psychotic drugs on the interaction of these two receptors.

Suggestions

Proteome-wide analysis of the role of expression of bacilysin operon on idiophase physiology of B. Subtilis
Demir, Mustafa; Özcengiz, Gülay; Department of Biology (2013)
The members of the genus Bacillus produce a wide variety of secondary metabolites with antimetabolic and pharmacological activities. These metabolites are mostly small peptides and have unusual components and chemical bonds. These metabolites are synthesized nonribosomally by multifunctional enzyme complexes called peptide synthetases. One of those small peptides, bacilysin, is a dipeptide antibiotic composed of L-alanine and L-anticapsin which is produced and excreted by certain strains of Bacillus subtili...
Comparative analysis of product and by-product distributions in defined and complex media in serine alkaline protease production by recombinant Basillus subtilis
Oktar, Ceren; Çalık, Pınar; Department of Chemical Engineering (2003)
In this study, firstly the effects of aspartic acid group amino acids -which were reported to be the potential bottleneck in serine alkaline protease (SAP) synthesis- on SAP production were investigated by substituting at a concentration range of 0-15 mM by using recombinant Bacillus subtilis carrying pHV1434::subC gene. All aspartic acid group amino acids except threonine inhibited SAP activity when CAA= 2.5 mM. The highest SAP activities with asparagine, aspartic acid, lysine, threonine, isoleucine and me...
Characterization of extracellular beta-lactamases from penicillin G-resistant cells of Streptococcus thermophilus
Chirica, LC; Güray, Nülüfer Tülün; Gültekin, Güzin Candan; Bozoglu, F (International Association for Food Protection, 1998-07-01)
In this study, biochemical properties of two extracellular beta-lactamases produced by penicillin-resistant Streptococcus thermophilus cells were investigated. Both beta-lactamases showed specificity for penicillins but not for cephaloridins. The p-lactamases exhibited different affinities for penicillin G. The one with the higher molecular weight (F1) had a K(m) value of 3.44 mu M and a V(max), value of 8.33, mu mol/min/mg of protein, whereas the beta-lactamase with the lower molecular weight (FII) had a K...
The effect of arginine-428 mutation on modulation of activity of human liver flavin monooxygenase 3 (FMO3) by imipramine and chlorpromazine
Adalı, Orhan; Philpot, RM (1999-07-01)
This study was carried out to investigate the molecular basis for modulation of recombinant FMO3-catalyzed activity by the tricyclicantidepressants, imipramine and chlorpromazine. A mutant of human liver FMO3 (T428R) was formed by site-directed mutagenesis and characterized along with the native enzyme in order to elucidate a possible structure-function relationship. Functional properties of native and T428R human FMO3s were studied with methimazole as substrate. Both enzymes catalyzed the S-oxidation of me...
A Comparative Study for the Evaluation of Two Doses of Ellagic Acid on Hepatic Drug Metabolizing and Antioxidant Enzymes in the Rat
Celik, Gurbet; SEMİZ, ASLI; Karakurt, Serdar; ARSLAN, ŞEVKİ; Adalı, Orhan; ŞEN, ALAATTİN (Hindawi Limited, 2013-01-01)
The present study was designed to evaluate different doses of ellagic acid (EA) in vivo in rats for its potential to modulate hepatic phases I, II, and antioxidant enzymes. EA (10 or 30 mg/kg/day, intragastrically) was administered for 14 consecutive days, and activity, protein, and mRNA levels were determined. Although the cytochrome P450 (CYP) 2B and CYP2E enzyme activities were decreased significantly, the activities of all other enzymes were unchanged with the 10 mg/kg/day EA. In addition, western-blot ...
Citation Formats
S. Akkuzu, “The Functional assessment of fluorescently tagged adenosine A2A and dopamine D2 receptors and qualitative analysis of dimerization of adenosine A2A and dopamine D2 receptor by using FRET,” M.S. - Master of Science, Middle East Technical University, 2013.