ER aminopeptidase 1 (ERAP1) customizes antigenic peptide precursors for MHC class I presentation and edits the repertoire. Coding single nucleotide polymorphisms (SNPs) in ERAP1 were recently linked with predisposition to autoimmune disease, suggesting a link between pathogenesis of autoimmunity ERAP1-mediated Ag processing. To investigate this possibility, we analyzed effect that disease-linked SNPs have on processing by vitro. Michaelis-Menten analysis revealed presence affects Michaelis...

The adipocyte‐derived leucine aminopeptidase (A‐LAP)/ER aminopeptidase‐1 is a multi‐functional enzyme belonging to the M1 family of aminopeptidases. It was reported that polymorphism Lys528Arg in human A‐LAP gene associated with essential hypertension. In this study, role Lys528 enzymatic activity examined by site‐directed mutagenesis. Among non‐synonymous polymorphisms tested, only reduced activity. replacement various amino acids including Ala, Met, His and Arg caused significant decrease...

Endoplasmic reticulum aminopeptidase 1 (ERAP1) is a multifunctional enzyme with an important role in processing antigenic peptides presented to class I major histocompatibility complex the endoplasmic reticulum. In this study, we found that reticulum-retained ERAP1 was secreted from macrophages response activation by treatment lipopolysaccharide (LPS) and interferon (IFN)-γ enhanced their phagocytic activity. Enhancement of activity murine macrophage RAW264.7 cells induced LPS/IFN-γ...

Perinatal treatment of female mice with natural and synthetic estrogens including diethylstilbestrol (DES) results in estrogen-independent persistent proliferation cornification the vaginal epithelium. The dynamics induction estrogen receptor (ER), c-jun, c-fos c-myc mRNAs by 17β-estradiol (E2) was examined uterus vagina neonatally DES-exposed -unexposed ovariectomized adult mice. In DES-unexposed mice, expression ER mRNA increased within 1 h after E2 administration declined 12 thereafter....

Aminopeptidase A (APA) is a type II membrane-bound protein implicated in the regulation of blood pressure brain renin-angiotensin system. In this study, recombinant soluble form APA was expressed baculovirus system, purified to homogeneity, and characterized. By using synthetic substrates, it shown that although enzyme has rather broad substrate specificity absence Ca2+, preferential release acidic amino acid residues observed presence Ca2+. Moreover, Ca2+ up- or down-regulated enzymatic...

Laeverin/aminopeptidase Q (APQ) is a cell surface protein specifically expressed on human embryo-derived extravillous trophoblasts that invades the uterus during placentation. The cDNA cloning of Laeverin/APQ revealed sequence encodes with 990 amino acid residues, and contains HEXXHX18E gluzincin motif, which characteristic M1 family aminopeptidases, although exopeptidase motif family, GAMEN, uniquely substituted for HAMEN sequence. In this study, we recombinant using baculovirus expression...

ERAP-1 (endoplasmic-reticulum aminopeptidase-1) is a multifunctional enzyme with roles in the regulation of blood pressure, angiogenesis and presentation antigens to MHC class I molecules. Whereas shows restricted specificity toward synthetic substrates, its substrate natural peptides rather broad. Because pathophysiological significance ERAP-1, it important elucidate molecular basis enzymatic action. In present study we used site-directed mutagenesis identify residues affecting human...

Abstract Macrophages play an important role in host defense under several immunological, inflammatory, and/or infectious conditions. In our previous work, we demonstrated that endoplasmic reticulum aminopeptidase 1 (ERAP1) was secreted from macrophages response to LPS and IFN-γ, it enhanced their phagocytic activity. this study, analyzed the mechanism of LPS/IFN-γ–induced ERAP1 secretion. secretion enzyme murine macrophage cell line RAW264.7 suppressed by polymyxin B. Several agonists TLRs,...

Journal Article Substrate-dependent nitric oxide synthesis by secreted endoplasmic reticulum aminopeptidase 1 in macrophages Get access Yoshikuni Goto, Goto * 1Faculty of Pharmaceutical Sciences, Teikyo-Heisei University, Nakano, Tokyo 164-8530, Japan; 2Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, 3Department Life School Agriculture, Meiji Kawasaki, Kanagawa 214-8571, and 4Department System Chemotherapy Molecular Graduate Kyoto 606-8501, Japan *Yoshikuni Faculty 4-21-2 Japan....

Endoplasmic reticulum aminopeptidase 1 (ERAP1) is a final trimming enzyme of N-extended antigenic peptides bound to major histocompatibility complex class I molecules in the endoplasmic (ER). In our previous work, we found that ERAP1 secreted from macrophages response activation by lipopolysaccharide and interferon-γ. this paper, searched for amino acid sequence protein important ER retention constructing chimeric proteins between 485 615 was significant. Moreover, comparing genomic...

Aminopeptidase A (APA) plays an important role in the regulation of blood pressure by mediating angiotensin II degradation renin-angiotensin system. The Ca2+-induced modulation enzymatic activity is most characteristic feature APA among M1 family aminopeptidases. In this study, we used site-directed mutagenesis for any residues responsible Ca2+ human APA. Alignment sequences members led to identification Asp-221 as a significant residue members. Replacement with Asn or Gln resulted loss...

Although perinatal exposure of female rats to estrogenic compounds produces irreversible changes in brain function, it is still unclear how the amount and timing those substances affect learning or if alters estrogen receptor α (ERα) expression hippocampus cortex. In adult rats, we investigated effects neonatal a model compound, ethinyl estradiol (EE), on passive avoidance ERα expression. Female Wistar-Imamichi were subcutaneously injected with oil, 0.02 mg/kg EE, 2 20 17β-estradiol within...

Extracellular vesicles (EVs) have been isolated from various sources, including primary and cultured cell lines body fluids. Previous studies, those conducted in our laboratory, reported the stability of EVs under storage conditions. human whole saliva were separated via size-exclusion chromatography. To simulate effects gastric or intestinal fluids on EVs, pepsin pancreatin was added to samples. Additionally, determine effect bile acids, sodium cholate added. The samples then subjected...

Abstract RNase L is responsible for the 2‐5A host defense system, an RNA degradation pathway present in cells of higher vertebrates that functions both antiviral and anticellular activities interferon. The activity tightly regulated exerted only presence 2‐5A. postulated mechanism its regulation as follows: N‐terminal half ankyrin‐repeat domain masks C‐terminal nuclease absence On binding at domain, forms a homodimer removes from to become active form. A conformational change key step this...

Human laeverin/aminopeptidase Q (LVRN/APQ) is a novel member of the M1 family zinc aminopeptidases and specifically expressed on cell surface human extravillous trophoblasts. Multiple sequence alignment aminopeptidase revealed that first Gly residue within conserved exopeptidase motif family, GXMEN motif, uniquely substituted for His in LVRN/APQ. In this study, we evaluated roles nonconserved His(379), comprising enzymatic properties We substitution His(379) with caused significant changes...

Extracellular vesicles (EVs), which are found in almost all cells and human body fluids, currently being studied as a source of pathophysiological information. Previously, we demonstrated that at least two types EVs can be isolated from whole saliva (WS) using enzymatic activity dipeptidyl peptidase IV (DPP IV) marker for differentiating the EV subsets. In present study, fractions, termed EV-I 20 k-ppt EV-II 100 k-ppt, were prepared by combination size-exclusion chromatography improved...

We have isolated brefeldin A (BFA)-resistant cell lines, KB/BF-1 and KB/BF-2, from the human epidermoid carcinoma KB line.The BFA-resistant phenotypes been stably maintained for more than 3 months in absence of BFA.KB/BF-1 KB/BF-2 showed 10-30-fold higher resistance to cytotoxicity BFA but were 2-3-fold sensitive monensin nigericin, cells.KB/BF-1 aberrant structures Golgi complex with poorly developed cisternae surrounded by many small vesicles.Immunocytochemical studies done antibodies...

Endoplasmic reticulum aminopeptidase 1 (ERAP1) is a multi-functional enzyme. In this study, we analysed its role in lipopolysaccharide-induced inflammatory response wild-type and ERAP1-knockout mice. Following lipopolysaccharide injection, ERAP1 was secreted into the blood, increasing leucine activity NO synthesis therein. Among amino acids tested, arginine concentration significantly increased mice compared to These results suggest that behaves similar acute-phase proteins, which are blood...

Aminopeptidase B (APB, EC 3.4.11.6) preferentially hydrolyzes the N-terminal basic amino acids of synthetic and peptide substrates requires a physiological concentration NaCl for optimal activity. In this study, we used site-directed mutagenesis molecular modeling to search an acid residue that is critical enzymatic properties human APB. Substitution Phe297 with Tyr caused significant decrease in hydrolytic activity toward as well chloride anion sensitivity. Molecular suggests contributes...

Human laeverin/aminopeptidase Q (APQ) is a novel member of the M1 family zinc aminopeptidases and specifically expressed on cell surface extravillous trophoblasts. In this study, we examined significance Gln-238 laeverin/APQ, putative S1 site residue, by site-directed mutagenesis for its enzymatic activity substrate specificity. Replacement with Ala caused significant change in specificity rather than decrease activity. These results indicate that important laeverin/APQ. addition, our data...