Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:Q86TM3 (cage)
 29,987 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Metals are thought to play a role in the structure of many viruses. The crystal structure of the T=3 icosahedral cowpea chlorotic mottle virus (CCMV) suggests the presence of 180 unique metal-binding sites in the assembled protein cage. Each of these sites is thought to involve the coordination of the metal by five amino acids contributed from two adjacent coat protein subunits. We have used fluorescence resonance energy transfer (FRET), from tryptophan residues proximal to the putative metal-binding sites, to probe Tb(III) binding to the virus. Binding of Tb(III) was investigated on the wild-type virus and a mutant where the RNA binding ability of the virus was removed. Tb(III) binding was observed both in the wild-type virus ( K(d)=19 microM) and the mutant ( K(d)=17 microM), as monitored by the increase in Tb(III) fluorescence (545 nm) and concomitant decrease in tryptophan fluorescence (342 nm). Competitive binding experiments showed Ca(II) to have about 100-fold less affinity for the binding sites ( K(d)=1.97 mM). This is the first direct evidence of metal binding to the putative metal-binding sites, originally suggested from the crystal structure of CCMV.
 ...
PMID:Metal binding to cowpea chlorotic mottle virus using terbium(III) fluorescence. 1450 76

The structure of monoamine oxidase B revealed three aromatic amino acid residues within contact distance of the flavin cofactor and a large number of aromatic residues in the substrate binding site. Circular dichroism (CD) spectroscopy can detect alterations in the environment of aromatic residues as a result of ligand binding or redox changes. CD spectra of MAO A indicate that a small inhibitor such d-amphetamine perturbs the aromatic residues very little, but binding of the larger pirlindole (2,3,3a,4,5,6-hexahydro-8-methyl-1H-pyrazino[3,2,1-j,k]carbazole hydrochloride) causes spectral changes consistent with the alteration of the environment of tyrosine and tryptophan residues in particular. Reduction of the flavin cofactor induces large enhancement of the CD signals in the aromatic region (260-310 nm). When covalent modification of the flavin by clorgyline accompanies reduction, the perturbation is even greater. In contrast to the static picture offered by crystallography, this study reveals changes in the aromatic cage on ligand binding and suggests that reduction of the cofactor substantially alters the environment of aromatic residues presumably near the flavin. In addition, the covalently modified reduced MAO A shows significant differences from the substrate-reduced enzyme.
 ...
PMID:Conformational changes in monoamine oxidase A in response to ligand binding or reduction. 1505 94

One manner to study the role of serotonin (5-HT) in behavioral functions is through nutritional manipulation of its precursor tryptophan (TRP). By means of the method of acute TRP depletion, plasma TRP levels can be reduced in a reversible way in both humans and rats. In the present study a TRP-free protein-carbohydrate mixture was used to investigate the behavioral effects of lowering TRP and 5-HT concentrations in adult male rats. These animals were tested in models of anxiety (open field, home cage emergence test), depression (forced swimming test) and cognition (object recognition test and Morris water escape test). The TRP-free protein-carbohydrate mixture substantially reduced the ratio TRP/SigmaLNAA within 2 and 4 h by 75 and 60%, respectively. It was found that 4 h after administration, the treatment did not affect anxiety-related behavior nor did it cause depressive-like behavior. Also, no treatment effect was found on spatial learning performance in a Morris water escape test. On the other hand, performance in an object recognition test was clearly impaired after TRP depletion. Taken together, these data suggest that acute lowered central 5-HT levels are not associated with changes in affective behavior (i.e. anxiety and depression), but do impair object memory in adult rats.
 ...
PMID:Acute tryptophan depletion induced by a gelatin-based mixture impairs object memory but not affective behavior and spatial learning in the rat. 1508 21

Tandem mass spectrometry (MS/MS) utilizing both electron capture dissociation (ECD) and collisionally activated dissociation (CAD) was used to develop a qualitative and quantitative analytical method for chiral analysis of individual amino acid residues in polypeptides. ECD produced a more distinct chiral recognition than CAD, which is attributed to the smaller degree of vibrational excitation in ECD. Several peptide and protein model systems were used in this study, including the smallest known protein, tryptophan cage, a lactoferrin peptide, and the biologically relevant opioid peptide, dermorphin. An adaptation of the kinetic method was used to quantify the degree of separation between fragmentation patterns of stereoisomeric peptides as a function of fragment ion abundances. The obtained calibration scale for relative abundances of d-amino acids in diastereomeric peptide mixtures was accurate to 1% for ECD and to 3-5% for CAD. It was found that separation and quantification of stereoisomers could be advantageously performed by nanoflow reversed-phase liquid chromatography, with the objective of on-line MS/MS limited to stereoisomer identification. This technique shows promise for the analysis of chiral substitution in peptides and proteins, broadening the application area for tandem mass spectrometry.
 ...
PMID:Distinguishing and quantifying peptides and proteins containing D-amino acids by tandem mass spectrometry. 1601 75

The role of polypeptide collapse and formation of intermediates in protein folding is still under debate. Miniproteins, small globular peptide structures, serve as ideal model systems to study the basic principles that govern folding. Experimental investigations of folding dynamics of such small systems, however, turn out to be challenging, because requirements for high temporal and spatial resolution have to be met simultaneously. Here, we demonstrate how selective quenching of an extrinsic fluorescent label by the amino acid tryptophan (Trp) can be used to probe folding dynamics of Trp-cage (TC), the smallest protein known to date. Using fluorescence correlation spectroscopy, we monitor folding transitions as well as conformational flexibility in the denatured state of the 20-residue protein under thermodynamic equilibrium conditions with nanosecond time resolution. Besides microsecond folding kinetics, we reveal hierarchical folding of TC, hidden to previous experimental studies. We show that specific collapse of the peptide to a molten globule-like intermediate enhances folding efficiency considerably. A single point mutation destabilizes the intermediate, switching the protein to two-state folding behavior and slowing down the folding process. Our results underscore the importance of preformed structure in the denatured state for folding of even the smallest globular structures. A unique method emerges for monitoring conformational dynamics and ultrafast folding events of polypeptides at the nanometer scale.
 ...
PMID:A microscopic view of miniprotein folding: enhanced folding efficiency through formation of an intermediate. 1626 42

Tea consumption in many cases is the main source of caffeine intake in humans. In the present study neurochemical and behavioural effects of long-term tea intake are monitored in rats. Long-term tea administration did not alter plasma tryptophan (TRP) but significantly attenuated brain TRP and 5-hydroxytryptamine (5-HT, serotonin) levels. Brain 5-hydroxyindole acetic acid (5-HIAA) was comparable in both tea-treated and control rats. An increase in home cage activity was observed after one week in rats taking tea as sole source of liquid, whereas no change on the activity was observed in an open field. Caffeinism has been associated with depression. The decreases of brain monoamine metabolism observed in present study are discussed as lowering of mood observed in tea or coffee consumers.
 ...
PMID:Neurochemical and behavioural effects of long term intake of tea. 1641 19

We investigate different protocols of the basin hopping technique for de novo protein folding. Using the protein free-energy force field PFF01 we report the reproducible all-atom folding of the 20-amino-acid tryptophan-cage protein [Protein Data Bank (PDB) code: 112y] and of the recently discovered 26-amino-acid potassium channel blocker (PDB code: 1wqc), which exhibits an unusual fold. We find that simulations with increasing cycle length and random starting temperatures perform best in comparison with other parametrizations. The basin hopping technique emerges as a simple but very efficient and robust workhorse for all-atom protein folding.
 ...
PMID:Basin hopping simulations for all-atom protein folding. 1646 Jan 93

Peripheral immune activation results in physiological and behavioral responses including changes in the level of behavioral arousal. One mechanism through which immune activation can influence these responses is via actions on brainstem neuromodulatory systems, including serotonergic systems. To investigate the effects of peripheral immune activation on serotonergic systems and behavior, and the potential role of prostanoids in mediating these effects, we compared the effects of intraperitoneal injections of lipopolysaccharide (LPS), in the presence or absence of the cyclooxygenase inhibitor indomethacin, on total plasma L-tryptophan concentrations, Fos expression in subdivisions of the brainstem raphe complex, and home cage behaviors. Peripheral LPS administration had no effect on total plasma L-tryptophan concentrations but increased Fos expression in serotonergic neurons selectively within the interfascicular (DRI) and ventrolateral (DRVL) subdivisions of the dorsal raphe nucleus 4 h following treatment; pretreatment with indomethacin blocked the LPS-induced increases in Fos expression within the DRI and DRVL. Peripheral LPS administration decreased measures of behavioral arousal including locomotion, rearing, climbing, and self-grooming; LPS administration had no effect on these behaviors in mice pretreated with indomethacin. The indomethacin-sensitive effects of LPS on Fos expression in the DRI may be due to selective activation of Type II serotonergic neurons which are largely restricted to the DRI region and have unique afferent regulatory mechanisms and behavioral correlates. Further studies of the effects of peripheral immune activation on DRI serotonergic systems may lead to a better understanding of the relationships among immune function, serotonergic systems, and behavior.
 ...
PMID:Lipopolysaccharide has indomethacin-sensitive actions on Fos expression in topographically organized subpopulations of serotonergic neurons. 1655 44

Kynurenic acid (KYNA) is a tryptophan metabolite synthesized and released by glia and recently shown to be a non-competitive antagonist of alpha7 nicotinic acetylcholine receptors at physiologically relevant concentrations, and NMDA receptors at higher concentrations. KYNA concentration is elevated in individuals with schizophrenia and those with Alzheimer's disease, two populations exhibiting cholinergic-related cognitive impairments. The present study investigated the effects of elevated KYNA concentration on conditioned stimulus processing in rats. For the first 2 days of the experiment, a subset of rats received intracerebroventricular infusions of either KYNA (0.1 microM) or vehicle and were either returned to the home cage or received non-reinforced presentations of a visual stimulus. All rats subsequently received presentations of the same visual stimulus followed by food reward during a 6-day training phase. In vehicle-treated rats, pre-exposure to the visual stimulus reduced orienting behaviour to the light (standing on the hind legs and orienting towards the visual stimulus) when it was later reinforced (i.e., conditioned orienting). In contrast, pre-exposure to the visual cue or 2 days of KYNA pretreatment reduced conditioned orienting behaviour. Finally, the reduction of orienting in KYNA-treated rats following pre-exposure was not as robust as in vehicle-treated rats. These results suggest that elevated KYNA levels can alter specific aspects of attentional processing of environmental stimuli and are discussed in terms of the potential contribution of KYNA to cognitive function and dysfunction.
 ...
PMID:Increased concentration of cerebral kynurenic acid alters stimulus processing and conditioned responding. 1662 Oct 49

Xenon-129 biosensors offer an attractive alternative to conventional MRI contrast agents due to the chemical shift sensitivity and large nuclear magnetic signal of hyperpolarized (129)Xe. Here, we report the first enzyme-responsive (129)Xe NMR biosensor. This compound was synthesized in 13 steps by attaching the consensus peptide substrate for matrix metalloproteinase-7 (MMP-7), an enzyme that is upregulated in many cancers, to the xenon-binding organic cage, cryptophane-A. The final coupling step was achieved on solid support in 80-92% yield via a copper (I)-catalyzed [3+2] cycloaddition. In vitro enzymatic cleavage assays were monitored by HPLC and fluorescence spectroscopy. The biosensor was determined to be an excellent substrate for MMP-7 (K(M) = 43 microM, V(max) = 1.3 x 10(-)(8) M s(-1), k(cat)/K(M) = 7,200 M(-1) s(-1)). Enzymatic cleavage of the tryptophan-containing peptide led to a dramatic decrease in Trp fluorescence, lambda(max) = 358 nm. Stern-Volmer analysis gave an association constant of 9000 +/- 1,000 M(-1) at 298 K between the cage and Trp-containing hexapeptide under enzymatic assay conditions. Most promisingly, (129)Xe NMR spectroscopy distinguished between the intact and cleaved biosensors with a 0.5 ppm difference in chemical shift. This difference most likely reflected a change in the electrostatic environment of (129)Xe, caused by the cleavage of three positively charged residues from the C-terminus. This work provides guidelines for the design and application of new enzyme-responsive (129)Xe NMR biosensors.
 ...
PMID:Designing 129Xe NMR biosensors for matrix metalloproteinase detection. 1701 9


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>