ABSTRACT
In the endoplasmic reticulum glycoprotein quality control system, UDP-glucose : glycoprotein glucosyltransferase (UGGT) functions as a folding sensor. Although it is known to form a heterodimer with selenoprotein F (SelenoF), the details of the complex formation remain obscure. A pulldown assay using co-transfected SelenoF and truncated mutants of human UGGT1 (HUGT1) revealed that SelenoF binds to the TRXL2 domain of HUGT1. Additionally, a newly developed photoaffinity crosslinker was selectively introduced into cysteine residues of recombinant SelenoF to determine the spatial orientation of SelenoF to HUGT1. The crosslinking experiments showed that SelenoF formed a covalent bond with amino acids in the TRXL3 region and the interdomain between ßS2 and GT24 of HUGT1 via the synthetic crosslinker. SelenoF might play a role in assessing and refining the disulfide bonds of misfolded glycoproteins in the hydrophobic cavity of HUGT1 as it binds to the highly flexible region of HUGT1 to reach its long hydrophobic cavity. Clarification of the SelenoF-binding domain of UGGT and its relative position will help predict and reveal the function of SelenoF from a structural perspective.
Subject(s)
Glucosyltransferases , Glycoproteins , Humans , Glucosyltransferases/metabolism , Glycoproteins/metabolism , Uridine Diphosphate , Selenoproteins , Glucose/metabolism , Protein FoldingABSTRACT
This study investigated the position of ultrasonic irradiation source and reactor geometry on fragmentation rate of a layered compound, α-zirconium phosphate (α-ZrP). By numerically solving the acoustic pressure distribution using COMSOL Multiphysics®, it is clarified the mechanism whereby the operating factors influenced the α-ZrP dispersion to make a suggestion of guideline of the process design method. Two vessels made of glass with a flat-bottom and a spherical-bottom, respectively, were used. Although the flat-bottom vessel at lower horn position showed the best performance of fragmentation, the region of high acoustic pressure field in the flat bottom vessel sharply narrowed and the transmittance became prominently low. On the other hand, no significant difference of the transmittance value in the spherical bottom vessel between the cases of low and high horn positions could be observed and the spherical bottom vessel was robust for the horn position. These results suggest that not only the magnitude of acoustic pressure but also the size of high acoustic pressure region is also an important factor and a spherical bottom vessel is one of suitable shape which gives large size of high acoustic pressure region regardless of the horn position.
ABSTRACT
Volatile organic compounds (VOCs) are considered the main cause of sick building syndrome and they are likely to irritate the skin, eyes, and mucous membrane; however, the toxic threshold and the mechanisms of cutaneous reaction induced by long-time VOC exposure have not been clarified. In the present study, we investigated the effect of repeated painting of VOCs onto mouse skin. Various concentrations of toluene, xylene, and formaldehyde (FA) were applied once a week for 5 weeks. While FA solution (2-10%) induced remarkable ear swelling and caused evident infiltration of inflammatory cells, high concentrations of toluene and xylene (50 or 100%) evoked mild ear swelling and marginal inflammatory cell invasion. In addition, FA exposure markedly increased the expression of interleukin-4 (IL-4), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and transient receptor potential vanilloid-1 (TRPV-1) mRNAs in the ears and IL-4 and NT-3 mRNAs in the cervical lymph nodes. Furthermore, capsazepine, a TRPV-1 antagonist, significantly suppressed ear swelling caused by repeated painting of 5% FA. These findings demonstrate that FA has more potent irritancy against skin than toluene or xylene and suggest that the Th2 response, neurotrophins and TRPV-1 play important roles in FA-induced skin inflammation.
