title: Monocytic elastase-mediated apolipoprotein-E degradation: Potential involvement of microglial elastase-like proteases in apolipoprotein-E proteolysis in brains with Alzheimers disease.
abstract: Impaired clearance of soluble Aβ (amyloid-β) promotes Aβ aggregation in brains with Alzheimer's disease (AD), while apolipoprotein-E (ApoE) in microglia mediates Aβ clearance. We studied the protease responsible for ApoE(4) degradation in human peripheral monocyte extracts, which are from the same lineage as microglia. We detected the hydrolytic activity for ApoE(4) in high-salt extracts with 2 M NaCl and found that the activity was inhibited by a serine protease inhibitor and an elastase-specific inhibitor, but not by other protease inhibitors. The extracts exhibited higher activity for the elastase substrate, and we followed the activity with ion-exchange and gel-filtration chromatography. Through silver staining, we partially purified a protein of 28 kDa, which was clarified as elastase by liquid chromatography-tandem mass spectrometry. These observations suggest that elastase is the key protease for ApoE(4) degradation. We also detected ApoE(4) hydrolytic activity in high-salt extracts in mouse microglial (BV-2) cell lysates, and showed that the ApoE(4) fragments by the BV-2 extracts differed from the fragments by the monocyte extracts. Though the ApoE(4) degradation by the extracts was not inhibited with elastase-specific inhibitors, it was inhibited by an elastase-specific monoclonal antibody, suggesting that elastase-like proteases in microglia differ from those of monocytes. Immunohistochemistry revealed that both elastase and ApoE were expressed in the senile plaques of brains with AD. In vitro studies also disclosed the localization of elastase in the microglial cell line, BV-2. Our results suggest that elastase-like proteases in the microglial cells surrounding Aβ plaques are responsible for ApoE degradation in the brain.
authors.affiliation: Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Nishihama, Yamashiro-cho, Tokushima 770-8514, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Suenaga
authors.affiliation: Department of Cellular and Molecular Neuropathology, Juntendo University, School of Medicine, Hongo 2-1-1, Bunkyou-ku, Tokyo 113-8421, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Furuta
authors.affiliation: Department of Neuropathology, Institute of Brain Science, Hirosaki University, Graduate School of Medicine, 5 Zaifu-cho, Hirosaki 036-8562, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Wakabayashi
authors.affiliation: Department of Gastroenterology and Hepatology, Kochi Medical School, Kochi University, Nankoku, Kochi 783-8505, Japan.
authors.forename: Toshiji
authors.initials: T
authors.lastname: Saibara
authors.affiliation: Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Nishihama, Yamashiro-cho, Tokushima 770-8514, Japan. Electronic address: matsunaga@ph.bunri-u.ac.jp.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Matsunaga
mesh_terms: D000368:Aged; D000544:Alzheimer Disease / Q000378:metabolism* / Q000473:pathology; D053327:Apolipoprotein E4 / Q000378:metabolism*; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D019272:Leukocyte Elastase / Q000378:metabolism*; D008297:Male; D017628:Microglia / Q000378:metabolism* / Q000473:pathology; D009000:Monocytes / Q000201:enzymology / Q000473:pathology; D059748:Proteolysis
chemical_list: D053327:Apolipoprotein E4; D019272:Leukocyte Elastase
keywords: Alzheimer's disease; Apolipoprotein-E; Elastase; Microglia; Protease