RESUMO
HYPOTHESIS: We have proposed a spreading-flow model of the solvent movement in a gel, which states that the solvent flows along the gel network segment much easier than in any other direction. An excess amount of the solvent component adsorbed on a flat solid surface in a gel will be transferred over the macroscopic gel network by the spreading-flow. Then, by measuring the concentration distribution of each solvent near the solid surface at the sub-millimeter scale, it should be possible to evaluate adsorption occurring on the nanometer scale. EXPERIMENTS: Confocal Raman microscopy was employed to map the concentration distributions of mixed solvents of p-xylene, mesitylene, and octanenitrile gelled by 12-hydroxystearic acid near the atomically flat mica surface. The measured concentration profiles were analyzed by the spreading-flow theory to construct adsorption isotherms. FINDINGS: Nearly all isotherms follow power laws with similar exponents, which is consistent with the van der Waals multilayer adsorption with weak adsorbate-adsorbent interactions. The present study demonstrates the concept of gel network amplification and its usefulness for probing nano-phenomena using macro-technology.
RESUMO
UDP-Glucuronosyltransferase (UGT) plays an important role in the metabolism of endogenous and exogenous compounds. UGT is a type I membrane protein, and has a dilysine motif (KKXX/KXKXX) in its C-terminal cytoplasmic domain. Although a dilysine motif is defined as an endoplasmic reticulum (ER) retrieval signal, it remains a matter of debate whether this motif functions in the ER localization of UGT. To address this issue, we generated systematic deletion mutants of UGT2B7, a major human isoform, and compared their subcellular localizations with that of an ER marker protein calnexin (CNX), using subcellular fractionation and immunofluorescent microscopy. We found that although the dilysine motif functioned as the ER retention signal in a chimera that replaced the cytoplasmic domain of CD4 with that of UGT2B7, UGT2B7 truncated mutants lacking this motif extensively colocalized with CNX, indicating dilysine motif-independent ER retention of UGT2B7. Moreover, deletion of the C-terminal transmembrane and cytoplasmic domains did not affect ER localization of UGT2B7, suggesting that the signal necessary for ER retention of UGT2B7 is present in its luminal domain. Serial deletions of the luminal domain, however, did not affect the ER retention of the mutants. Further, a cytoplasmic and transmembrane domain-deleted mutant of UGT2B7 was localized to the ER without being secreted. These results suggest that UGT2B7 could localize to the ER without any retention signal, and lead to the conclusion that the static localization of UGT results from lack of a signal for export from the ER.
