The N terminus of the human alpha(1D)-adrenergic receptor prevents cell surface expression

We previously reported that truncation of the N-terminal 79 amino acids of alpha(1D)-adrenoceptors (Delta(1-79)alpha(1D)-ARs) greatly increases binding site density. In this study, we determined whether this effect was associated with changes in alpha(1D)-AR subcellular localization. Confocal imaging of green fluorescent protein (GFP)-tagged receptors and sucrose density gradient fractionation suggested that full-length alpha(1D)-ARs were found primarily in intracellular compartments, whereas Delta(1-79)alpha(1D)-ARs were translocated to the plasma membrane. This resulted in a 3- to 4-fold increase in intrinsic activity for stimulation of inositol phosphate formation by norepinephrine. We determined whether this effect was transplantable by creating N-terminal chimeras of alpha(1)-ARs containing the body of one subtype and the N terminus of another (alpha(1A) NT-D, alpha(1B) NT-D, alpha(1D) NT-A, and alpha(1D)NT-B). When expressed in human embryonic kidney 293 cells, radioligand binding revealed that binding densities of alpha(1A)- or alpha(1B)-ARs containing the alpha(1D)-N terminus decreased by 86 to 93%, whereas substitution of alpha(1A)- or alpha(1B)-N termini increased alpha(1D)-AR binding site density by 2- to 3-fold. Confocal microscopy showed that GFP-tagged alpha(1D)NT-B-ARs were found only on the cell surface, whereas GFP-tagged alpha(1B)NT-D-ARs were completely intracellular. Radioligand binding and confocal imaging of GFP-tagged alpha(1D)- and Delta(1-79)alpha(1D)-ARs expressed in rat aortic smooth muscle cells produced similar results, suggesting these effects are generalizable to cell types that endogenously express alpha(1D)-ARs. These findings demonstrate that the N-terminal region of alpha(1D)-ARs contain a transplantable signal that is critical for regulating formation of functional bindings, through regulating cellular localization.

Laser-flash photolysis of alpha-brominated o-xylenes

The flash photolysis (lambda = 266 nm) of four alpha -brominated omicron -xylenes in apolar solvents gives two transients characterized, depending on parent compound substitution, as either monoradicals or carbenes, and quinodimethanes. alpha -Bromomethylbenzyl radical is characterized and the novel species alpha,alpha'-dibromo-omicron -xylylene and omicron-(alpha,alpha -dibromomethyl)benzyl carbene are described for the first time. (C) 2001 Elsevier Science B.V. All rights reserved.

Muscle delta C-13 change in Nile tilapia (Oreochromis niloticus): Effects of growth and carbon turnover

The contribution of growth and turnover to the muscle delta C-13 change process was investigated using mathematical models which associate delta C-13 change to time of intake of a new diet or increase in body mass. Two groups of Nile tilapia (Oreochromis niloticus) were fed on diets based on C3 (sigma C-13 = - 25.64 +/- 0.06 parts per thousand) or C4 (delta C-13= -16.01 +/- 0.06 parts per thousand) photosynthetic cycle plants to standardize the muscle delta C-13. After establishing the carbon isotopic equilibrium, fish (mean mass 24.12 +/- 6.79 g) then received the other treatment diet until a new carbon isotopic equilibrium could be established, characterizing T1 (C3-C4) and T2 (C4-C3) treatments. No significant differences were observed in fish productive performance. Good fits were obtained for the models that associated the delta C-13 change to time, resulting in carbon half-life values of 23.33 days for T1 and 25.96 days for T2. Based on values found for the muscle delta C-13 change rate from growth (0.0263 day(-1) and 0.0254 day(-1)) and turnover (0.0034 day(-1) and 0.0013 day(-1)), our results indicate that most of the delta C-13 change could be attributed to growth. The application of model that associated the delta C-13 change to body mass increase seems to produce results with no apparent biological explanation. The delta C-13 change rate could directly reflect the daily ration and growth rate, and consequently the isotopic change rates of carbon and other tissue elements can be properly used to assess different factors that may interfere in nutrient utilization and growth. (c) 2006 Published by Elsevier B.V.

A stability transition at mildly acidic pH in the alpha-hemolysin (alpha-toxin) from Staphylococcus aureus

The effects of mildly acidic conditions on the free energy of unfolding (Delta G(u)(buff)) of the pore-forming alpha-hemolysin (alpha HL) from Staphylococcus aureus were assessed between pH 5.0 and 7.5 by measuring intrinsic tryptophan fluorescence, circular dichroism and elution time in size exclusion chromatography during urea denaturation, Decreasing the pH from 7.0 to 5.0 reduced the calculated Delta G(u)(buff) from 8.9 to 4.2 kcal moI(-1), which correlates with an increased rate of pore formation previously observed over the same pH range, It is proposed that the lowered surface pH of biological membranes reduces the stability of alpha HL thereby modulating the rate of pore formation. (C) 1999 Federation of European Biochemical Societies.

Structure, organization, and expression of the alpha prolamin multigenic family bring new insights into the evolutionary relationships among grasses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)