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Organic & Biomolecular Chemistry
Page 4 of 5
DOI: 10.1039/C7OB02391J
COMMUNICATION
Journal Name
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Figure 5. FRRIAR-TUC alone and in the presence of apo CaM resulted in no hp 129Xe
NMR signal (a); CaM bound to FRRIAR-TUC resulted in two distinct 129Xe NMR
resonances (b). NMR performed in 10 mM HEPES, 1 mM CaCl2, pH 7.2 with 1% DMSO.
129Xeaq peak shown at 192.8 ppm. CaM binding peptide structure taken from PDB 1SY9.
Image generated in PyMOL (Delano Scientific, LLC; South San Francisco, CA USA).
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In conclusion, we report a straightforward synthesis for the
first “turn-on” cryptophane-xenon biosensor, which was used
to detect Ca2+-bound CaM protein. This so-called turn-on
effect improves signal-to-noise, especially in scenarios where
there are multiple biosensors with both “free” and “bound”
signals. The observation of signal only in the presence of target
analyte will also facilitate ultrasensitive detection using
hyperpolarized chemical exchange saturation transfer (Hyper-
CEST) NMR techniques.16e For example, large-spectral-width,
high-power pulses that increase detection sensitivity can be
employed without interfering signal from untargeted
biosensor(s). Thus, this work highlights a potential new
paradigm for cryptophane-based xenon biosensors. Future
studies will help to elucidate the nature of peptide-
cryptophane interactions that can produce the “OFF” state.
This research was supported by National Institutes of
Health (NIH R01-GM097478). EJP acknowledges support for
CaM projects through the National Science Foundation (NSF
CHE-1150351). We thank Dr. Rakesh Kohli for assistance with
MALDI-MS (NSF MRI-0820996). We thank Drs. George Furst
and Jun Gu for assistance with NMR spectroscopy. We also
thank the University of Pennsylvania Biological Chemistry
Resource Center for automated peptide synthesis supported
by UPenn Laboratory for Research on the Structure of Matter
(NSF MRSEC DMR 0520020, 1120901). RFW thanks the NIH for
funding through the Chemistry-Biology Interface Training
Program (T32 GM07133). We thank Dr. Jacob Goldberg for
help with data analysis.
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4 | J. Name., 2012, 00, 1-3
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