Cytochromeâ P450â Induced Ordering of Microsomal Membranes Modulates Affinity for Drugs

Although membrane environment is known to boost drug metabolism by mammalian cytochromeâ P450s, the factors that stabilize the structural folding and enhance protein function are unclear. In this study, we use peptideâ based lipid nanodiscs to â trapâ the lipid boundaries of microsomal cytochromeâ P450 2B4. We report the first evidence that CYP2B4 is able to induce the formation of raft domains in a biomimetic compound of the endoplasmic reticulum. NMR experiments were used to identify and quantitatively determine the lipids present in nanodiscs. A combination of biophysical experiments and molecular dynamics simulations revealed a sphingomyelin binding region in CYP2B4. The proteinâ induced lipid raft formation increased the thermal stability of P450 and dramatically altered ligand binding kinetics of the hydrophilic ligand BHT. These results unveil membrane/protein dynamics that contribute to the delicate mechanism of redox catalysis in lipid membrane.Redox catalysis in the lipid membrane: A novel application of peptide nanodiscs shows that cytochromeâ P450 2B4 is able to induce the formation of lipid raft domains in a biomimetic compound of the endoplasmic reticulum (ER). The proteinâ induced lipid rafts increase the thermal stability cytochromeâ P450 and dramatically alter the ligandâ binding kinetics of the hydrophilic ligand BHT.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/142960/1/anie201713167.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/142960/2/anie201713167_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/142960/3/anie201713167-sup-0001-misc_information.pd

J. agric. food chem.

Removal of basal leaves near blooms inevitably affects grapevine balance and cluster microclimate conditions, improving fruit quality. Mechanization of this practice allows growers to save time and resources, but to our knowledge, it has not yet been compared with the manual application of this practice in a cool-climate region where seasonal temperatures frequently limit fruit technological maturity and phenolic ripening in red Vitis vinifera cultivars. In our research, berry sugar concentration was highest with prebloom mechanical treatment (PB-ME). Furthermore, metabolomics analysis revealed that PB-ME favored the accumulation of significantly more disubstituted anthocyanins and flavonols and OH-substituted anthocyanins compared with manual application. Given that vine balance was similar between treatments, increased ripening with PB-ME is likely due to enhanced microclimate conditions and higher carbon partitioning through a younger canopy containing basal leaf fragments proximal to fruit. This information provides an important strategy for consistently ripening red Vitis vinifera cultivars in cool climates

Increase in seed tannin extractability and oxidation using a freeze-thaw treatment in cool-climate grown red (Vitis vinifera L.) cultivars

Grape seed maturation involves the gradual oxidation of tannins, decreasing excessive bitterness and astringency in wine. In cool climates, this process is limited by the short growing season, affecting wine quality. A “freeze-thaw” treatment on seeds of red vinifera cultivars at veraison and harvest was used to evaluate the effect of oxidation and extractability on seed phenolic fractions. Freezing increased the extraction of total phenolics and o-diphenols quantified from fractionation (fraction 1, vacuolar tannins; fraction 2, hydrogen bonded tannins; fraction 3, covalently bonded tannins), especially at harvest. Despite this, colorimetry, microscopy, oxidation reactivity index (ORI), and correlations between the color index and fractions indicated that freezing disrupted vacuole integrity, enhancing oxidation in the seed coat. In conclusion, vacuolar tannins (which are the main seed phenolics extracted during fermentation) were highly correlated with seed color change, potentially providing information for winemaking in cool climate regions