TY - JOUR
T1 - Identification of Protein Targets of 12/15-Lipoxygenase-Derived Lipid Electrophiles in Mouse Peritoneal Macrophages Using Omega-Alkynyl Fatty Acid
AU - Isobe, Yosuke
AU - Kawashima, Yusuke
AU - Ishihara, Tomoaki
AU - Watanabe, Kenji
AU - Ohara, Osamu
AU - Arita, Makoto
N1 - Funding Information:
This work was funded in part by the Japan Society for the Promotion of Science KAKENHI JP15H05897, 15H05898, and 15H04648 (M.A.) and the RIKEN Special Postdoctoral Researcher Program (Y.I.). We thank Yuki Ariyasu for skillful technical assistance in immunocytochemistry.
Publisher Copyright:
© 2018 American Chemical Society.
PY - 2018/4/20
Y1 - 2018/4/20
N2 - The 12/15-lipoxygenase (12/15-LOX) enzyme introduces peroxyl groups, in a position-specific manner, into polyunsaturated fatty acids to form various kinds of bioactive lipid metabolites, including lipid-derived electrophiles (LDE). The resident peritoneal macrophage is the site of highest 12/15-LOX expression in the mouse. However, the role of the enzyme in the regulation of resident macrophages is not fully understood. Here, we describe a chemoproteomic method to identify the targets of enzymatically generated LDE. By treating mouse peritoneal macrophages with omega-alkynyl arachidonic acid (aAA), we identified a series of proteins adducted by LDE generated through a 12/15-LOX catalyzed reaction. Pathway analysis revealed a dramatic enrichment of proteins involved in energy metabolism and found that glycolytic flux and mitochondrial respiration were significantly affected by the expression of 12/15-LOX. Our findings thus highlight the utility of chemoproteomics using aAA for identifying intracellular targets of enzymatically generated LDE.
AB - The 12/15-lipoxygenase (12/15-LOX) enzyme introduces peroxyl groups, in a position-specific manner, into polyunsaturated fatty acids to form various kinds of bioactive lipid metabolites, including lipid-derived electrophiles (LDE). The resident peritoneal macrophage is the site of highest 12/15-LOX expression in the mouse. However, the role of the enzyme in the regulation of resident macrophages is not fully understood. Here, we describe a chemoproteomic method to identify the targets of enzymatically generated LDE. By treating mouse peritoneal macrophages with omega-alkynyl arachidonic acid (aAA), we identified a series of proteins adducted by LDE generated through a 12/15-LOX catalyzed reaction. Pathway analysis revealed a dramatic enrichment of proteins involved in energy metabolism and found that glycolytic flux and mitochondrial respiration were significantly affected by the expression of 12/15-LOX. Our findings thus highlight the utility of chemoproteomics using aAA for identifying intracellular targets of enzymatically generated LDE.
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U2 - 10.1021/acschembio.7b01092
DO - 10.1021/acschembio.7b01092
M3 - Article
C2 - 29461797
AN - SCOPUS:85045852004
SN - 1554-8929
VL - 13
SP - 887
EP - 893
JO - ACS chemical biology
JF - ACS chemical biology
IS - 4
ER -