title: Substrate-dependent nitric oxide synthesis by secreted endoplasmic reticulum aminopeptidase 1 in macrophages.
abstract: In this study, we examined the role of aminopeptidases with reference to endoplasmic reticulum aminopeptidase 1 (ERAP1) in nitric oxide (NO) synthesis employing murine macrophage cell line RAW264.7 cells activated by lipopolysaccharide (LPS) and interferon (IFN)-γ and LPS-activated peritoneal macrophages derived from ERAP1 knockout mouse. When NO synthesis was measured in the presence of peptides having N-terminal Arg, comparative NO synthesis was seen with that measured in the presence of Arg. In the presence of an aminopeptidase inhibitor amastatin, NO synthesis in activated RAW264.7 cells was significantly decreased. These results suggest that aminopeptidases are involved in the NO synthesis in activated RAW264.7 cells. Subsequently, significant reduction of NO synthesis was observed in ERAP1 knockdown cells compared with wild-type cells. This reduction was rescued by exogenously added ERAP1. Furthermore, when peritoneal macrophages prepared from ERAP1 knockout mouse were employed, reduction of NO synthesis in knockout mouse macrophages was also attributable to ERAP1. In the presence of amastatin, further reduction was observed in knockout mouse-derived macrophages. Taken together, these results suggest that several aminopeptidases play important roles in the maximum synthesis of NO in activated macrophages in a substrate peptide-dependent manner and ERAP1 is one of the aminopeptidases involved in the NO synthesis.
authors.affiliation: Faculty of Pharmaceutical Sciences, Teikyo-Heisei University, Nakano, Tokyo 164-8530, Japan; Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan; Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan; and Department of System Chemotherapy and Molecular Sciences, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan y.goto@thu.ac.jp.
authors.forename: Yoshikuni
authors.initials: Y
authors.lastname: Goto
authors.affiliation: Faculty of Pharmaceutical Sciences, Teikyo-Heisei University, Nakano, Tokyo 164-8530, Japan; Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan; Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan; and Department of System Chemotherapy and Molecular Sciences, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Ogawa
authors.affiliation: Faculty of Pharmaceutical Sciences, Teikyo-Heisei University, Nakano, Tokyo 164-8530, Japan; Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan; Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan; and Department of System Chemotherapy and Molecular Sciences, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan Faculty of Pharmaceutical Sciences, Teikyo-Heisei University, Nakano, Tokyo 164-8530, Japan; Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan; Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan; and Department of System Chemotherapy and Molecular Sciences, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Takahiro J
authors.initials: TJ
authors.lastname: Nakamura
authors.affiliation: Faculty of Pharmaceutical Sciences, Teikyo-Heisei University, Nakano, Tokyo 164-8530, Japan; Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan; Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan; and Department of System Chemotherapy and Molecular Sciences, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Hattori
authors.affiliation: Faculty of Pharmaceutical Sciences, Teikyo-Heisei University, Nakano, Tokyo 164-8530, Japan; Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan; Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan; and Department of System Chemotherapy and Molecular Sciences, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Tsujimoto
mesh_terms: D000626:Aminopeptidases / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D008264:Macrophages / Q000201:enzymology*; D051379:Mice; D018345:Mice, Knockout; D015778:Minor Histocompatibility Antigens; D009569:Nitric Oxide / Q000096:biosynthesis*; D013379:Substrate Specificity
chemical_list: D015778:Minor Histocompatibility Antigens; D009569:Nitric Oxide; D000626:Aminopeptidases; C509944:ERAP1 protein, mouse
keywords: aminopeptidase; arginine; lipopolysaccharide; macrophage; nitric oxide