RRC ID 54806
Author Aki T, Unuma K, Noritake K, Hirayama N, Funakoshi T, Uemura K.
Title Formation of high molecular weight p62 by CORM-3.
Journal PLoS One
Abstract CORM-3 is a water-soluble carbon monoxide (CO)-releasing molecule developed for possible therapeutic use of CO. CORM-3 belongs to a group of metal carbonyl compounds that contain transition metals and carbonyls as the central scaffold and coordinated ligands, respectively. CORM-3 has been reported to be reactive with many proteins in eukaryotes including mammals. Among them, several extracellular proteins, such as lysozyme, as well as plasma albumin and fibronectin, have been shown to interact directly with CORM-3. p62 is an intracellular adaptor protein required for targeting ubiquitinated (Ub) proteins to lysosomal degradation through autophagy. p62 has been shown to undergo self-oligomerization via covalent crosslinking in response to treatment with verteporfin, a benzoporphyrin derivative used for photodynamic therapy. Here we show that CORM-3 also interacts directly with p62. When applied to mouse embryonic fibroblasts (MEFs) at a high concentration (1 mM), CORM-3 causes the formation of reduction- and detergent-resistant high molecular weight (HMW)-p62. HMW-p62 accumulates more in atg5-/- MEFs than in wild type (WT) MEFs, showing the elimination of HMW-p62 through autophagy. HMW-p62 is also generated in H9c2 rat cardiomyoblastoma as well as A549 human alveolar epithelial cells, suggesting that HMW-p62 formation is not specific to MEFs, but, rather, is a general event in mammalian cells. HMW-p62 formation by CORM-3 can be reproduced using purified p62 in vitro, demonstrating the direct interaction between CORM-3 and p62. These results show that p62 is a CORM-3-interactive intracellular protein.
Volume 14(1)
Pages e0210474
Published 2019-1-8
DOI 10.1371/journal.pone.0210474
PII PONE-D-18-27112
PMID 30620762
PMC PMC6324786
MeSH A549 Cells Animals Apoptosis / drug effects Autophagy-Related Protein 5 / deficiency Autophagy-Related Protein 5 / metabolism Cattle Fibroblasts / cytology Fibroblasts / drug effects Fibroblasts / metabolism Gene Knockdown Techniques Humans Mice Molecular Weight Organometallic Compounds / pharmacology* Protein Aggregates RNA, Small Interfering / metabolism RNA-Binding Proteins / chemistry* RNA-Binding Proteins / metabolism Rats Ubiquitin / metabolism Verteporfin / pharmacology
IF 2.776
Times Cited 1
Human and Animal Cells Atg5^(+/+)MEF(RCB2710) Atg5^(-/-)MEF(RCB2711) A549(RCB0098)