TY - JOUR
T1 - μmap-Red
T2 - Proximity Labeling by Red Light Photocatalysis
AU - Buksh, Benito F.
AU - Knutson, Steve D.
AU - Oakley, James V.
AU - Bissonnette, Noah B.
AU - Oblinsky, Daniel G.
AU - Schwoerer, Michael P.
AU - Seath, Ciaran P.
AU - Geri, Jacob B.
AU - Rodriguez-Rivera, Frances P.
AU - Parker, Dann L.
AU - Scholes, Gregory D.
AU - Ploss, Alexander
AU - Macmillan, David W.C.
N1 - Funding Information:
This work was funded by the NIH National Institute of General Medical Sciences (R35-GM134897-02) and kind gifts from Merck, BMS, Pfizer, Janssen, Genentech, and Eli Lilly. We also acknowledge the Princeton Catalysis Initiative for supporting this work. S.D.K. acknowledges the NIH for a postdoctoral fellowship (1F32GM142206-01). J.V.O. acknowledges the National Science Foundation Graduate Research Fellowship Program (DGE-1656466). Mechanistic experiments, including transient absorption and UV–vis spectroscopy, were supported by the Division of Chemical Sciences, Geosciences, and Biosciences, Office of Basic Energy Sciences of the U.S. Department of Energy (DOE) through grant DE-SC0019370. M.P.S. is supported by the NIGMS of the National Institutes of Health under Grant T32GM007388 and a HMEI-STEP fellowship from the from the High Meadows Environmental Institute at Princeton University. Imaging was performed with support from the Confocal Imaging Facility, a Nikon Center of Excellence, in the Department of Molecular Biology at Princeton University. The authors thank Saw Kyin and Henry H. Shwe at the Princeton Proteomics Facility. We thank Christina DeCoste, Katherine Rittenbach, and the Princeton University Molecular Biology Flow Cytometry Resource Facility which is partially supported by the Cancer Institute of New Jersey Cancer Center Support Grant (P30CA072720). Generalized schemes were created using Biorender.
Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/4/13
Y1 - 2022/4/13
N2 - Modern proximity labeling techniques have enabled significant advances in understanding biomolecular interactions. However, current tools primarily utilize activation modes that are incompatible with complex biological environments, limiting our ability to interrogate cell-and tissue-level microenvironments in animal models. Here, we report μMap-Red, a proximity labeling platform that uses a red-light-excited SnIVchlorin e6 catalyst to activate a phenyl azide biotin probe. We validate μMap-Red by demonstrating photonically controlled protein labeling in vitro through several layers of tissue, and we then apply our platform in cellulo to label EGFR microenvironments and validate performance with STED microscopy and quantitative proteomics. Finally, to demonstrate labeling in a complex biological sample, we deploy μMap-Red in whole mouse blood to profile erythrocyte cell-surface proteins. This work represents a significant methodological advance toward light-based proximity labeling in complex tissue environments and animal models.
AB - Modern proximity labeling techniques have enabled significant advances in understanding biomolecular interactions. However, current tools primarily utilize activation modes that are incompatible with complex biological environments, limiting our ability to interrogate cell-and tissue-level microenvironments in animal models. Here, we report μMap-Red, a proximity labeling platform that uses a red-light-excited SnIVchlorin e6 catalyst to activate a phenyl azide biotin probe. We validate μMap-Red by demonstrating photonically controlled protein labeling in vitro through several layers of tissue, and we then apply our platform in cellulo to label EGFR microenvironments and validate performance with STED microscopy and quantitative proteomics. Finally, to demonstrate labeling in a complex biological sample, we deploy μMap-Red in whole mouse blood to profile erythrocyte cell-surface proteins. This work represents a significant methodological advance toward light-based proximity labeling in complex tissue environments and animal models.
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U2 - 10.1021/jacs.2c01384
DO - 10.1021/jacs.2c01384
M3 - Article
C2 - 35363468
AN - SCOPUS:85128219845
SN - 0002-7863
VL - 144
SP - 6154
EP - 6162
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 14
ER -