Journal of the American Chemical Society
Communication
incorporated into a cellular lipid pool and utilized as an acyl
donor in palmitoylation.
(10) Saleem, A. N.; Chen, Y. H.; Baek, H. J.; Hsiao, Y. W.; Huang, H.
W.; Kao, H. J.; Liu, K. M.; Shen, L. F.; Song, I. W.; Tu, C. P.; Wu, J. Y.;
Kikuchi, T.; Justice, M. J.; Yen, J. J.; Chen, Y. T. PLoS Genet. 2010, 6,
No. e1000985.
In conclusion, we have synthesized and characterized activity-
based probes 2 and 3 based on 1, a known irreversible pan-
inhibitor of PATs. As DHHC-PATs are among the proteins
labeled by the probes, 2 and 3 could be used to facilitate the
identification of additional palmitoylating enzymes as well as in
proteomic analyses of PATs in signaling, normal physiology,
and diseases. In addition, they will also provide a format for
identifying inhibitors of PATs for therapeutic applications. It is
noted that DHHC-PATs are not the only cellular targets of 1
and its analogues. Indeed, probe 2 labeled many non-
palmitoylating enzymes in our mass spectrometry studies.
Because of the high reactivity, off-target effects, and non-
selectivity, many classes of activity-based probes often label
proteins outside of their intended target class. For example, a
probe designed to target protein kinases also labels many
(
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2
(
Hoogendoorn, S.; van der Marel, G. A.; Florea, B. I.; Overkleeft, H. S.
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33
nonkinases in mass spectrometry studies. Although the
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specificity of ABPP probes is not absolute, they are valuable
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15
for enriching and characterizing disease-relevant enzymes.
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M. L.; Deschenes, R. J.; Linder, M. E. J. Lipid Res. 2009, 50, 233.
The future development of additional activity-based probes
with improved specificity and efficiency for targeting PATs
might address the limitations of the probes described here.
(
(
(
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ASSOCIATED CONTENT
Supporting Information
Synthesis, experimental protocols, supporting figures, full list of
identified proteins from mass spectrometry studies (XLS) and
■
(22) Jennings, B. C.; Linder, M. E. J. Biol. Chem. 2012, 287, 7236.
(23) Chase, J. F.; Tubbs, P. K. Biochem. J. 1972, 129, 55.
(24) Noritake, J.; Fukata, Y.; Iwanaga, T.; Hosomi, N.; Tsutsumi, R.;
Matsuda, N.; Tani, H.; Iwanari, H.; Mochizuki, Y.; Kodama, T.;
Matsuura, Y.; Bredt, D. S.; Hamakubo, T.; Fukata, M. J. Cell Biol. 2009,
*
S
1
86, 147.
(25) Martin, B. R.; Cravatt, B. F. Nat. Methods 2009, 6, 135.
(
26) Charron, G.; Zhang, M. M.; Yount, J. S.; Wilson, J.; Raghavan,
AUTHOR INFORMATION
A. S.; Shamir, E.; Hang, H. C. J. Am. Chem. Soc. 2009, 131, 4967.
(27) Hannoush, R. N.; Arenas-Ramirez, N. ACS Chem. Biol. 2009, 4,
5
(
81.
28) Martin, B. R.; Wang, C.; Adibekian, A.; Tully, S. E.; Cravatt, B.
F. Nat. Methods 2012, 9, 84.
29) Yang, W.; Di Vizio, D.; Kirchner, M.; Steen, H.; Freeman, M. R.
Notes
The authors declare no competing financial interest.
(
Mol. Cell. Proteomics 2010, 9, 54.
ACKNOWLEDGMENTS
(30) Ducker, C. E.; Griffel, L. K.; Smith, R. A.; Keller, S. N.; Zhuang,
Y.; Xia, Z.; Diller, J. D.; Smith, C. D. Mol. Cancer Ther. 2006, 5, 1647.
■
We thank Ross Tomaino at the Taplin Mass Spectrometry
Facility of Harvard Medical School for proteomics studies and
Drs. Matthew Tremblay and Jianming Zhang for critical
comments on the manuscript. This work was supported by
the Massachusetts General Hospital (MGH) Institutional Fund
and the Stewart Rahr-MRA Young Investigator Award (X.W.).
(
31) Bachovchin, D. A.; Brown, S. J.; Rosen, H.; Cravatt, B. F. Nat.
Biotechnol. 2009, 27, 387.
32) Yang, Y. Y.; Grammel, M.; Raghavan, A. S.; Charron, G.; Hang,
(
H. C. Chem. Biol. 2010, 17, 1212.
(33) Patricelli, M. P.; Szardenings, A. K.; Liyanage, M.; Nomanbhoy,
T. K.; Wu, M.; Weissig, H.; Aban, A.; Chun, D.; Tanner, S.; Kozarich,
J. W. Biochemistry 2007, 46, 350.
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