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. 2020 May 19;jbc.RA120.013867.
doi: 10.1074/jbc.RA120.013867. Online ahead of print.

Autophagy Induction by Exogenous Polyamines Is an Artifact of Bovine Serum Amine Oxidase Activity in Culture Serum

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Autophagy Induction by Exogenous Polyamines Is an Artifact of Bovine Serum Amine Oxidase Activity in Culture Serum

Cassandra E Holbert et al. J Biol Chem. .
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Abstract

Polyamines are small polycationic alkylamines involved in many fundamental cellular processes, including proliferation, nucleic acid synthesis, apoptosis, and protection from oxidative damage. It has been proposed that in addition to these functions, elevated levels of polyamines promote longevity in various biological systems, including yeast, drosophila, and murine models. A series of in vitro mechanistic studies by multiple investigators has led to the conclusion that addition of exogenous spermidine promotes longevity through autophagy induction; however, these experiments were confounded by the use of mammalian cell culture systems supplemented with fetal bovine serum (FBS). Using cell viability assays, LC3B immunoblots, and live-cell fluorescence microscopy, we report here that in the presence of ruminant serum, exogenously added polyamines are quickly oxidized by the copper-containing bovine serum amine oxidase (BSAO). This polyamine oxidation resulted in the production of harmful byproducts including hydrogen peroxide, ammonia, and reactive aldehydes. Our data demonstrate that it is critically important to prevent confounding BSAO-induced cytotoxicity in mechanistic studies of the roles of polyamines in autophagy.

Keywords: amine oxidase; autophagy; beta-oxidation; bovine serum amine oxidase; cytotoxicity; hydrogen peroxide; oxidative stress; polyamine; spermidine; spermine.

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