The metabolism of arsenic is generally accepted to proceed by repetitive reduction and oxidative methylation; the latter is mediated by arsenic methyltransferase (Cyt19). In human urine, the major metabolites of inorganic arsenicals such as arsenite (iAsIII) and arsenate (iAsV) are monomethylarsonic acid (MMA(V)) and dimethylarsinic acid (DMA(V)). On the other hand, in rat bile, the major metabolites of iAsIII have been reported to be arsenic-glutathione (As-GSH) complexes. In the present study we investigate whether these As-GSH complexes are substrates for arsenic methyltransferase by using human recombinant Cyt19. Analyses by high-performance liquid chromatography-inductively coupled plasma mass spectrometry suggested that arsenic triglutathione (ATG) was generated nonenzymatically from iAsIII when GSH was present at concentrations 2 mM or higher. Human recombinant Cyt19 catalyzed transfer of a methyl group from S-adenosyl-L-methionine to arsenic and produced monomethyl and dimethyl arsenicals. The methylation of arsenic was catalyzed by Cyt19 only when ATG was present in the reaction mixture. Moreover, monomethylarsonic diglutathione (MADG) was a substrate of Cyt19 for further methylation to dimethylarsinic glutathione (DMAG). On the other hand, monomethylarsonous acid (MMA(III)), a hydrolysis product of MADG, was not methylated to dimethyl arsenical by Cyt19. These results suggest that As-GSH complexes such as ATG and MADG were converted by Cyt19 to MADG and DMAG, respectively. Both MADG and DMAG were unstable in solution when the GSH concentration was lower than 1 mM, and were hydrolyzed and oxidized to MMA(V) and DMA(V), respectively. Metabolism of iAsIII to methylated arsenicals by Cyt19 was via ATG and MADG rather than by oxidative methylation of iAsIII and MMA(III).

Moore, G. F., Taira, A., Klaus, A., Becker, L., Boeckel, B., Cragg, B. A., Dean, A., Fergusson, C. L., Henry, P., Hirano, S., Hisamitsu, T. et al. (2001). New insights into deformation and fluid flow processes in the Nankai Trough accretionary prism: Results of Ocean Drilling Program Leg 190. Geochemistry, Geophysics, Geosystems, 2, Article No: 2001GC000166.The Nankai Trough accretionary prism is considered an ?end-member? prism accreting a coarse terrigenous sediment section in a setting with structural simplicity, unparalleled resolution by seismic and other geophysical techniques, and large historic earthquakes. It therefore has been the focus of Ocean Drilling Program (ODP) drilling to address several unresolved questions concerning accretionary processes and prism evolution. At six sites cored along two transects across the Nankai Trough accretionary prism during ODP Leg 190, lithostratigraphy and sediment diagenesis vary markedly. For the first time, reference sites at the seaward ends of the two transects defined the stratigraphic framework of the accreting/subducting Shikoku Basin sedimentary section. A thick section of Miocene turbidites and smectite-rich mudstone is present within the subducting section at the Ashizuri site. The turbidites and mudstones are absent in the correlative section at the Muroto site; variations in lithology, mineralogy, and hydrologic properties of the incoming sediments probably contribute to the difference in prism wedge taper between the two transects, while possibly controlling the seismic character of the active plate boundary. The d?collement in both transects is localized within a common stratigraphic unit (?5.9?7 Ma) within the lower Shikoku Basin facies. The d?collement is also a major boundary for both physical and mechanical properties. A broad low-chloride pore water anomaly in the lower Shikoku Basin unit, first identified at Site 808, progressively decreases in magnitude from prism to basin along the Muroto Transect. Physical properties relationships, evidence for mineralogic changes in the sediments, and pore fluid chemistry suggest that the chloride anomaly results primarily from in situ diagenetic reactions in the sediments, possibly augmented by flow of freshened fluid from depth. New constraints on stratigraphy and age of units along more landward parts of the Muroto Transect have dramatically changed our ideas about the tectonic evolution of the prism in this area. Growth of the seaward-most part of the prism took place very rapidly, with 40 km of accretion within the past 2 Myr. This rate is at least 3 times greater than growth rates in a comparable prism.Peer reviewe

For the past three decades, most attention in heavy metal toxicology has been paid to cadmium, mercury, lead, chromium, nickel, vanadium, and tin because these metals widely polluted the environment. However, with the development of new materials in the last decade, the need for toxicological studies on those new materials has been increasing. (Suppl 11:85-95 (1996)

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Ubiquitination serves as a key sorting signal in the lysosomal degradation of endocytosed receptors through the ability of ubiquitinated membrane proteins to be recognized and sorted by ubiquitin-binding proteins along the endocytic route. The ESCRT-II complex in yeast contains one such protein, Vps36, which harbors a ubiquitin-binding NZF domain and is required for vacuolar sorting of ubiquitinated membrane proteins. Surprisingly, the presumptive mammalian ortholog Eap45 lacks the ubiquitin-binding module of Vps36, and it is thus not clear whether mammalian ESCRT-II functions to bind ubiquitinated cargo. In this paper, we provide evidence that Eap45 contains a novel ubiquitinbinding domain, GLUE (GRAM-like ubiquitin-binding in Eap45), which binds ubiquitin with similar affinity and specificity as other ubiquitin-binding domains. The GLUE domain shares similarities in its primary and predicted secondary structures to phosphoinositide-binding GRAM and PH domains. Accordingly, we find that Eap45 binds to a subset of 3-phosphoinositides, suggesting that ubiquitin recognition could be coordinated with phosphoinositide binding. Furthermore, we show that Eap45 colocalizes with ubiquitinated proteins on late endosomes. These results are consistent with a role for Eap45 in endosomal sorting of ubiquitinated cargo.Ubiquitination of proteins was until recently mainly considered to be important for targeting proteins for degradation in the 26S proteasome (1, 2). The recent discovery that ubiquitination may work as a reversible modification has, however, shown that the attachment of ubiquitin is also essential for changes in protein location, activity, and interaction with binding partners. Ubiquitination is thus emerging as a very important mechanism for regulating several cellular processes, such as signal transduction, membrane trafficking, transcriptional regulation, virus budding, and DNA repair (3). Mono-and multi-ubiquitination have been reported to regulate the internalization of several membrane receptors, such as receptor tyrosine kinases and G-protein-coupled receptors, from the plasma membrane into clathrin-coated pits (4, 5). In addition, ubiquitination is required for sorting endocytosed membrane proteins destined for degradation into intraluminal vesicles of multivesicular bodies (MVBs).1 The MVBs eventually fuse with lysosomes to degrade the internal vesicles and their protein cargo (6, 7).One way cells interpret and transmit the mono-and multiubiquitination signal is through the action of proteins that bind ubiquitin noncovalently (3). Several proteins implicated in endocytosis and sorting of receptors have been found to contain a variety of ubiquitin binding domains, such as the ubiquitininteracting motif (UIM) (8, 9), the ubiquitin-associated domain (UBA) (10, 11), the Cue1-related domain (CUE) (12, 13), the ubiquitin E2 enzyme variant domain (UEV or Ubc-like) (14, 15), and the Npl4-zink finger (NZF) domain (16,17).Many of the subcomponents of the molecular machinery responsible for lysosomal sorting have...

Hrs has an essential role in sorting of monoubiquitinated receptors to multivesicular bodies for lysosomal degradation, through recognition of ubiquitinated receptors by its ubiquitin-interacting motif (UIM). Here, we present the structure of a complex of Hrs-UIM and ubiquitin at 1.7-A resolution. Hrs-UIM forms a single alpha-helix, which binds two ubiquitin molecules, one on either side. These two ubiquitin molecules are related by pseudo two-fold screw symmetry along the helical axis of the UIM, corresponding to a shift by two residues on the UIM helix. Both ubiquitin molecules interact with the UIM in the same manner, using the Ile44 surface, with equal binding affinities. Mutational experiments show that both binding sites of Hrs-UIM are required for efficient degradative protein sorting. Hrs-UIM belongs to a new subclass of double-sided UIMs, in contrast to its yeast homolog Vps27p, which has two tandem single-sided UIMs.

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