Claudin-21 Has a Paracellular Channel Role at Tight Junctions

Claudin protein family members, of which there are at least 27 in humans and mice, polymerize to form tight junctions (TJs) between epithelial cells, in a tissue- and developmental stage-specific manner. Claudins have a paracellular barrier function. In addition, certain claudins function as paracellular channels for small ions and/or solutes by forming selective pores at the TJs, although the specific claudins involved and their functional mechanisms are still in question. Here we show for the first time that claudin-21, which is more highly expressed in the embryonic than the postnatal stages, acts as a paracellular channel for small cations, such as Na⁺, similar to the typical channel-type claudins claudin-2 and -15. Claudin-21 also allows the paracellular passage of larger solutes. Our findings suggest that claudin-21-based TJs allow the passage of small and larger solutes by both paracellular channel-based and some additional mechanisms.     

Incorporation of glycosylated amino acid into protein by an in vitro translation system

O-GalNAcα-modified proteins are the precursor of mucin-type O-glycosylated proteins. Homogeneously O-glycosylated proteins are required to investigate the biological functions of glycoproteins and to develop biopharmaceuticals. Here we show that the incorporation of GalNAcα-Thr into proteins successfully proceeded by the use of a chemically aminoacylated tRNA. GalNAcα-Thr was chemoenzymatically attached to amber suppressor tRNA and the product was subjected to in vitro translation together with streptavidin mRNA containing the UAG codon. Gel electrophoresis and mass analysis showed that GalNAcα-Thr was successfully incorporated into the N-terminus, although it was not incorporated at the interior. This method will facilitate the preparation of homogeneous GalNAcα-proteins.     

3-Isoxazolidone from jack bean seedlings

3-Isoxazolidone has been isolated from jack bean seedlings.     

Studies on Morphological and Physio-Ecological Variations of the Reniform Nematode,Rotylenchulus reniformis Linford and Oliveira, 1940 with an Emphasis on Differential Geographical Distribution of Amphimictic and Parthenogenetic Populations in Japan

The geographical distribution and polymorphism in morphological and biological characters of the reniform nematode, Rotylenchulus reniformis, in Japan were studied. The northern limit of habitat of this nematode was found on the 14 °C isothermal line of annual average-air temperature. Three morphologically different groups were ascertained which corresponded to three biological types based on male frequency. Incidence of males was consistent within populations and was not affected by environmental factors. Sexual attraction of males by females indicated reproductive isolation between the male-numerous type (MNT) and male-rare type (MRT) or male-absent type (MAT). Reproduction was amphimictic in the MNT and parthenogenetic in the MRT and MAT. Larval development in both MRT and MAT, but not that of MNT popula-tions, was inhibited at 34 °C. Differences in host preference were also observed among populations.     

Probing the Transmembrane Structure and Dynamics of Microsomal NADPH-cytochrome P450 oxidoreductase by Solid-State NMR

NADPH-cytochrome P450 oxidoreductase (CYPOR) is an essential redox partner of the cytochrome P450 (cyt P450) superfamily of metabolic enzymes. In the endoplasmic reticulum of liver cells, such enzymes metabolize ∼75% of the pharmaceuticals in use today. It is known that the transmembrane domain of CYPOR plays a crucial role in aiding the formation of a complex between CYPOR and cyt P450. Here we present the transmembrane structure, topology, and dynamics of the FMN binding domain of CYPOR in a native membrane-like environment. Our solid-state NMR results reveal that the N-terminal transmembrane domain of CYPOR adopts an α-helical conformation in the lipid membrane environment. Most notably, we also show that the transmembrane helix is tilted ∼13° from the lipid bilayer normal, and exhibits motions on a submillisecond timescale including rotational diffusion of the whole helix and fluctuation of the helical director axis. The approaches and the information reported in this study would enable further investigations on the structure and dynamics of the full-length NADPH-cytochrome P450 oxidoreductase and its interaction with other membrane proteins in a membrane environment.