TY - JOUR
T1 - A Photoactivatable Formaldehyde Donor with Fluorescence Monitoring Reveals Threshold to Arrest Cell Migration
AU - Smaga, Lukas P.
AU - Pino, Nicholas W.
AU - Ibarra, Gabriela E.
AU - Krishnamurthy, Vishnu
AU - Chan, Jefferson
N1 - Funding Information:
This work was supported the National Institutes of Health (R35GM133581). N.W.P. and G.E.I. acknowledge the Chemistry–Biology Interface Training Grant (T32 GM070421) and the Alfred P. Sloan Foundation for financial support. N.W.P. thanks Dr. Robert C. and Mrs. Carolyn J. Springborn for a graduate fellowship. G.E.I. thanks the National Consortium for Graduate Degrees for Minorities in Engineering & Science for a graduate fellowship. Major funding for the 500 MHz Bruker CryoProbeTM was provided by the Roy J. Carver Charitable Trust (Muscatine, Iowa; Grant No. 15-4521) to the School of Chemical Sciences NMR Lab. The Q-Tof Ultima mass spectrometer was purchased in part with a grant from the National Science Foundation, Division of Biological Infrastructure (DBI-0100085). We also acknowledge the Core Facilities at the Carl R. Woese Institute for Genomic Biology for access to the Zeiss LSM 700 confocal microscope and corresponding software. We acknowledge Dr. Iwona Dobrucka and the Molecular Imaging Laboratory at the Beckman Institute for use of the IVIS imaging system. We thank Mr. Lucas D. Akin for aid with mass spectrometry experiments, Prof. Paul J. Hergenrother and Mr. Matthew Boudreau for assistance with apoptotic assays, Prof. Kai Zhang for help with construction of an LED array, and Ms. Ashley Chan for the design of the graphical abstract.
PY - 2020/1/15
Y1 - 2020/1/15
N2 - Controlled light-mediated delivery of biological analytes can enable the investigation of highly reactivity molecules within living systems. As many biological effects are concentration dependent, it is critical to determine the location, time, and quantity of analyte donation. In this work, we have developed the first photoactivatable donor for formaldehyde (FA). Our optimized photoactivatable donor, photoFAD-3, is equipped with a fluorescence readout that enables monitoring of FA release with a concomitant 139-fold fluorescence enhancement. Tuning of photostability and cellular retention enabled quantification of intracellular FA release through cell lysate calibration. Application of photoFAD-3 uncovered the concentration range necessary for arresting wound healing in live cells. This marks the first report where a photoactivatable donor for any analyte has been used to quantify intracellular release.
AB - Controlled light-mediated delivery of biological analytes can enable the investigation of highly reactivity molecules within living systems. As many biological effects are concentration dependent, it is critical to determine the location, time, and quantity of analyte donation. In this work, we have developed the first photoactivatable donor for formaldehyde (FA). Our optimized photoactivatable donor, photoFAD-3, is equipped with a fluorescence readout that enables monitoring of FA release with a concomitant 139-fold fluorescence enhancement. Tuning of photostability and cellular retention enabled quantification of intracellular FA release through cell lysate calibration. Application of photoFAD-3 uncovered the concentration range necessary for arresting wound healing in live cells. This marks the first report where a photoactivatable donor for any analyte has been used to quantify intracellular release.
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U2 - 10.1021/jacs.9b11899
DO - 10.1021/jacs.9b11899
M3 - Article
C2 - 31898899
AN - SCOPUS:85077935404
VL - 142
SP - 680
EP - 684
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 2
ER -