title: Protective effect of recombinant soluble neprilysin against β-amyloid induced neurotoxicity.
abstract: A few decades ago, researchers found emerging evidence showing that a number of sequential events lead to the pathological cascade of Alzheimer's disease (AD) which is caused by the accumulation of amyloid beta (Aβ), a physiological peptide, in the brain. Therefore, regulation of Aβ represents a crucial treatment approach for AD. Neprilysin (NEP), a membrane metallo-endopeptidase, is a rate-limiting peptidase which is known to degrade the amyloid beta peptide. This study investigated soluble NEP (sNEP) produced by recombinant mammalian cells stably transfected with a non-viral NEP expression vector to demonstrate its protective effect against Aβ peptides in neuronal cells in vitro. Stably transfected HEK 293 cells were used to purify the soluble protein. sNEP and Aβ peptide co-treated hippocampal cells had a decreased level of Aβ peptides shown by an increase in cell viability and decrease in apoptosis measured by the CCK-8 and relative caspase-3 activity ratio assays, respectively. This study shows that stably transfected mammalian cells can produce soluble NEP proteins which could be used to protect against Aβ accumulation in AD and subsequently neuronal toxicity. Additionally, approaches using protein therapy for potential targets could change the pathological cascade of Alzheimer's disease.
authors.affiliation: Department of Chemistry, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-747, Republic of Korea.
authors.forename: Jae Hoon
authors.initials: JH
authors.lastname: Oh
authors.affiliation: Department of Chemistry, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-747, Republic of Korea.
authors.forename: Sunghyun
authors.initials: S
authors.lastname: Choi
authors.affiliation: Department of Chemistry, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-747, Republic of Korea.
authors.forename: Jeehae
authors.initials: J
authors.lastname: Shin
authors.affiliation: Department of Chemistry, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-747, Republic of Korea. Electronic address: pfjspark@snu.ac.kr.
authors.forename: Jong-Sang
authors.initials: JS
authors.lastname: Park
mesh_terms: D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors* / Q000633:toxicity; D057809:HEK293 Cells; D006801:Humans; D015260:Neprilysin / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: D016229:Amyloid beta-Peptides; D011994:Recombinant Proteins; D015260:Neprilysin
keywords: Alzheimer’s disease; Amyloid beta peptide; Neurotoxicity; Protein theraphy; Recombinant soluble neprilysin