A5049356705- Cellular transport and secretion
- Lipid Membrane Structure and Behavior
- Endoplasmic Reticulum Stress and Disease
- Epigenetics and DNA Methylation
- Pancreatic function and diabetes
- Phagocytosis and Immune Regulation
- Calcium signaling and nucleotide metabolism
- Genetic Syndromes and Imprinting
- Receptor Mechanisms and Signaling
- Cancer Immunotherapy and Biomarkers
- Biotin and Related Studies
- Immune cells in cancer
- Ubiquitin and proteasome pathways
- Fungal and yeast genetics research
- Erythrocyte Function and Pathophysiology
- Bladder and Urothelial Cancer Treatments
- Advanced Proteomics Techniques and Applications
- Retinal Development and Disorders
- Prenatal Screening and Diagnostics
- Monoclonal and Polyclonal Antibodies Research
- RNA modifications and cancer
- Ion channel regulation and function
- RNA regulation and disease
- Fibroblast Growth Factor Research
- Toxin Mechanisms and Immunotoxins
Tottori University
2015-2024
Fukushima Medical University
2004-2018
Institute of Macromolecular Chemistry
2015
Institute of Biomedical Science
2005-2008
Japan Science and Technology Agency
2005-2008
The University of Tokyo
2000-2005
Tokyo University of Pharmacy and Life Sciences
1997-2004
Max Planck Society
2004
Max Planck Institute for Biophysical Chemistry
2003
University of Tsukuba
1989-1994
Many peptide hormones are produced from larger precursors by endoproteolysis at pairs of basic amino acids (e.g. Lys-Arg and Arg-Arg) within the regulated secretory pathway in endocrine cells. However, many other membrane proteins appear to be through cleavage multiple, rather than paired, residues constitutive non-endocrine By surveying various processed constitutively, we noticed that most them have consensus sequence, Arg-X-Lys/Arg-Arg (RXK/RR), site. When expressed cells, a precursor...
We have cloned and sequenced a mouse cDNA encoding the 793-residue amino acid sequence of furin, which is protein homologous to yeast Kex2 protease. The entire 94% identical that human it contains 289-residue subtilisin-like catalytic domain. Within this region, 99, 64, 53% acids are those PC2 (the other mammalian Kex2-like protein), Kex2, respectively. It has been proposed furin prohormone processing enzyme cleaves precursors at paired basic acids, based on fact protease responsible for...
Synaptic exocytosis requires the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins syntaxin 1, SNAP-25, and synaptobrevin (VAMP). Assembly of SNAREs into a stable core complex is supposed to catalyze membrane fusion, proteoliposomes reconstituted with synaptic SNARE spontaneously fuse each other. We now show that liposome fusion mediated by inhibited botulinum neurotoxin E (BoNT/E) but can be rescued supplementing C-terminal portion SNAP-25. Furthermore,...
Furin, a mammalian homolog of the yeast Kex2 protease, is associated with Golgi membranes and involved in cleavage precursor proteins at sites marked by Arg-X-Lys/Arg-Arg (RXK/RR) motif. We have recently shown that furin mutant lacking transmembrane domain can be secreted from cDNA-transfected cells proteolytic activity for fluorogenic peptide t-butoxycarbonyl-Arg-Val-Arg-Arg-4-methylcoumarin-7- amide. In this study, we purified characterized recombinant conditioned medium these cells. Furin...
Tomosyn is a 130-kDa syntaxin-binding protein that contains large N-terminal domain with WD40 repeats and C-terminal homologous to R-SNAREs. Here we show tomosyn forms genuine SNARE core complexes the SNAREs syntaxin 1 SNAP-25. In vitro studies recombinant proteins revealed complex formation proceeds from unstructured monomers stable four-helical bundle. The assembled displayed features typical for complexes, including profound hysteresis upon unfolding-refolding transitions. No were formed...
Proteolytic cleavage (nicking) of diphtheria toxin (DT) in the 14-amino acid loop subtended by disulfide bond between Cys186 and Cys201 is required for cytotoxic action DT. The includes consensus motif a membrane-anchored protease, furin. We found that soluble form furin cleaves intact DT Arg103 Ser194 vitro. LoVo cells, human colon carcinoma cell line, do not produce functional show here cleaved cells. cells are resistant to DT, although they sensitive nicked before it added medium. When...
The hepatocyte growth factor/scatter factor (HGF/SF) receptor consists of an α‐ and a β‐subunit, which are derived from single‐chain precursor by endoproteolytic processing. is not proteolytically processed in LoVo colon carcinoma cells. uncleaved immunopurified the cells was cleaved vitro furin. Furthermore, HGF/SF transfected with furin cDNA. These results indicate that processing endoprotease for receptor. Tyrosine autophosphorylation induced HGF/SF, expressing stimulated indicating...
Obesity has tremendous impact on the health systems. Its epigenetic bases are unclear. MacroH2A1 is a variant of histone H2A, present in two alternatively exon-spliced isoforms macroH2A1.1 and macroH2A1.2, regulating cell plasticity proliferation, during pluripotency tumorigenesis. Their role adipose tissue unknown. Here, we show evidence that protein levels visceral obese humans positively correlate with BMI, while macroH2A1.2 nearly absent. We thus introduced constitutive GFP-tagged...
Members of the syntaxin family are target-soluble<i>N</i>-ethylmaleimide-sensitive factor-attachment protein receptors involved in vesicle docking and/or fusion within exocytic and endocytotic pathways. By using yeast two-hybrid system, we have identified a novel member family, 18, that binds to α-soluble <i>N</i>-ethylmaleimide-sensitive protein. Subcellular fractionation immunocytochemical analysis revealed 18 is principally located endoplasmic reticulum. We examined effect overexpression...
Many peptide hormones and neuropeptides are produced from larger, inactive precursors through endoproteolysis at sites usually marked by paired basic residues (primarily Lys-Arg Arg-Arg), or occasionally a monobasic residue Arg). Based upon data concerning processing of prorenin its mutants around the native cleavage site expressed in mouse pituitary AtT-20 cells, we present following sequence rules that govern mono-arginyl cleavages: (a) fourth (position -4) sixth -6) upstream is required,...
We have recently shown that furin, a mammalian homologue of the yeast precursor-processing endoprotease Kex2, is involved in precursor cleavage at sites marked by Arg-X-Lys/Arg-Arg motif within constitutive secretory pathway. In this study, we analyzed molecular and enzymatic properties furin expressed Chinese hamster ovary cells using gene transfer techniques. COOH-terminal truncation analyses indicate polypeptide region significantly conserved among Kex2 family members required for...
We cloned and sequenced a cDNA from library of mouse pituitary AtT-20 cells which are known to cleave an endogenous various foreign prohormones at dibasic sites. This encodes novel 753-residue protein, named PCS, is structurally related the yeast Kex2 protease involved in precursor cleavage sites recently identified mammalian Kex2-like proteins, furin PC2. Among examined cell lines tissues, PC3 mRNA was only detected cells. The substrate specificity expressed similar that observed conclude...
The endoplasmic reticulum (ER) is thought to play an important structural and functional role in phagocytosis. According this model, direct membrane fusion between the ER plasma or phagosomal must precede further invagination, but exact mechanisms remain elusive. Here, we investigated whether various ER-localized SNARE proteins are involved process. When phagosomes were isolated from murine J774 macrophages, found that (syntaxin 18, D12, Sec22b) significantly enriched phagosomes....
Macrophages that express representative endoplasmic reticulum (ER) molecules tagged with green fluorescence protein were generated to assess the recruitment of ER Leishmania parasitophorous vacuoles (PVs). More than 90% PVs harbouring pifanoi or donovani parasites recruited calnexin, their PV membrane. An equivalent proportion also membrane-associated soluble N-ethylmaleimide-sensitive factor attachment receptors (SNAREs), Sec22b. Both appeared be very early in formation nascent PVs....
Molecular imaging employing fluorescent proteins has been widely used to highlight specific reactions or processes in various fields of the life sciences. Despite extensive improvements tag, this technology is still limited study molecular events extracellular milieu. This partly due presence cysteine proteins. These almost cotranslationally form disulfide bonded oligomers when expressed endoplasmic reticulum (ER). Although single molecule photobleaching analysis showed that these were not...
VCP/p97 is involved in a variety of cellular processes, including membrane fusion and ubiquitin-dependent protein degradation. It has been suggested that adaptor proteins such as p47 Ufd1p confer functional versatility to VCP/p97. To identify novel adaptors, we searched for interact specifically with by using the yeast two-hybrid system, discovered VCP/p97-interacting named smallVCP/p97-interactingprotein (SVIP). Rat SVIP 76-amino acid contains two putative coiled-coil regions, potential...
Abstract Background Programmed cell death 1 (PD-1) is one of the immune checkpoint molecules that negatively regulate function T cells. Although recent studies indicate PD-1 also expressed on other cells besides cells, its role remains unclear. This study aims to evaluate expression macrophages and examine effect anti-tumor immunity in gastric cancer (GC) patients. Methods The frequency + obtained from GC tissue was determined by multicolor flow cytometry ( n = 15). Double...
Membrane‐anchored Neuregulin β1 sheds its ectodomain as soluble factors. Two proteases that belong to a disintegrin and metalloprotease (ADAM) family are known cleave β1. One is tumor necrosis factor‐α converting enzyme (TACE/ADAM17). The other Meltrin β (ADAM19). Against our expectation shedding by ADAM occurs at the cell surface, here we found mediates of in Golgi apparatus. was localized around apparatus developing sensory neurons. Subcellular fractionation revealed generated...
Synaptosomal associated protein of 23 kDa (SNAP-23), a plasma membrane–localized soluble N-ethylmaleimide–sensitive factor attachment receptor (SNARE), has been implicated in phagocytosis by macrophages. For elucidation its precise role this process, macrophage line overexpressing monomeric Venus–tagged SNAP-23 was established. These cells showed enhanced Fc receptor–mediated phagocytosis. Detailed analyses each process revealed marked increase the production reactive oxygen species within...
Newly synthesized mammalian secretory proteins such as preprolactin are translocated across the endoplasmic reticulum (ER) in a signal recognition particle (SRP)-dependent manner. Recent studies revealed that there two steps for peptides during this translocation. The first step is by SRP, which results elongation arrest, and second interaction between translocation channel embedded ER membrane. To determine roles of hydrophobic region SRP membrane-embedded machinery, we constructed chimeric...
The endoplasmic reticulum (ER) is proposed to be a membrane donor for phagosome formation. In support of this, we have previously shown that the expression level syntaxin 18, an ER-localized SNARE protein, correlates with phagocytosis activity. To obtain further insights into involvement ER in focused on Sec22b, another protein also found phagosomal membranes. marked contrast effects report here was nearly abolished J774 macrophages stably expressing mVenus-tagged without affecting cell...