TY - JOUR
T1 - Comparative Photoaffinity Profiling of Omega-3 Signaling Lipid Probes Reveals Prostaglandin Reductase 1 as a Metabolic Hub in Human Macrophages
AU - Gagestein, Berend
AU - Von Hegedus, Johannes H.
AU - Kwekkeboom, Joanneke C.
AU - Heijink, Marieke
AU - Blomberg, Niek
AU - Van Der Wel, Tom
AU - Florea, Bogdan I.
AU - Van Den Elst, Hans
AU - Wals, Kim
AU - Overkleeft, Herman S.
AU - Giera, Martin
AU - Toes, René E.M.
AU - Ioan-Facsinay, Andreea
AU - Van Der Stelt, Mario
N1 - Funding Information: This work was supported by grant ICI-00016 from the Institute for Chemical Immunology. M.v.d.S. received base-funding from Oncode Institute. We thank Caprotec for kindly providing the Caprobox. Dr. Richard van den Berg is acknowledged for feedback on the manuscript. Publisher Copyright: © 2022 American Chemical Society. All rights reserved.
PY - 2022/10/19
Y1 - 2022/10/19
N2 - The fish oil constituent docosahexaenoic acid (DHA, 22:6 n-3) is a signaling lipid with anti-inflammatory properties. The molecular mechanisms underlying the biological effect of DHA are poorly understood. Here, we report the design, synthesis, and application of a complementary pair of bio-orthogonal, photoreactive probes based on the polyunsaturated scaffold DHA and its oxidative metabolite 17-hydroxydocosahexaenoic acid (17-HDHA). In these probes, an alkyne serves as a handle to introduce a fluorescent reporter group or a biotin-affinity tag via copper(I)-catalyzed azide-alkyne cycloaddition. This pair of chemical probes was used to map specific targets of the omega-3 signaling lipids in primary human macrophages. Prostaglandin reductase 1 (PTGR1) was identified as an interaction partner that metabolizes 17-oxo-DHA, an oxidative metabolite of 17-HDHA. 17-oxo-DHA reduced the formation of pro-inflammatory lipids 5-HETE and LTB4 in human macrophages and neutrophils. Our results demonstrate the potential of comparative photoaffinity protein profiling for the discovery of metabolic enzymes of bioactive lipids and highlight the power of chemical proteomics to uncover new biological insights.
AB - The fish oil constituent docosahexaenoic acid (DHA, 22:6 n-3) is a signaling lipid with anti-inflammatory properties. The molecular mechanisms underlying the biological effect of DHA are poorly understood. Here, we report the design, synthesis, and application of a complementary pair of bio-orthogonal, photoreactive probes based on the polyunsaturated scaffold DHA and its oxidative metabolite 17-hydroxydocosahexaenoic acid (17-HDHA). In these probes, an alkyne serves as a handle to introduce a fluorescent reporter group or a biotin-affinity tag via copper(I)-catalyzed azide-alkyne cycloaddition. This pair of chemical probes was used to map specific targets of the omega-3 signaling lipids in primary human macrophages. Prostaglandin reductase 1 (PTGR1) was identified as an interaction partner that metabolizes 17-oxo-DHA, an oxidative metabolite of 17-HDHA. 17-oxo-DHA reduced the formation of pro-inflammatory lipids 5-HETE and LTB4 in human macrophages and neutrophils. Our results demonstrate the potential of comparative photoaffinity protein profiling for the discovery of metabolic enzymes of bioactive lipids and highlight the power of chemical proteomics to uncover new biological insights.
UR - http://www.scopus.com/inward/record.url?scp=85139488853&partnerID=8YFLogxK
U2 - https://doi.org/10.1021/jacs.2c06827
DO - https://doi.org/10.1021/jacs.2c06827
M3 - Article
C2 - 36197299
SN - 0002-7863
VL - 144
SP - 18938
EP - 18947
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 41
ER -