Journal of the American Chemical Society
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Schneckenburger, H.; Weber, P.; Wagner, M.; Schickinger,
that steric hindrance between the phenyl substituent and the
indandione moiety in P-Mero4 and its derivatives causes it to
adopt the unusual S-cis conformer. This is the first time that a
stable S-cis conformation is observed in fluorescent dyes. Be-
sides being sensitive to polarity and H-bonding, the dye also
exhibits strong fluorescence in viscous solvents and crowded
surroundings due to the restricted one-bond rotation of the
phenyl substituent on the tetramethine chain. This environ-
ment-sensitive feature of P-Mero4 was exploited to generate
fluorogenic probes, NAPH-Mero4SBG, P-Mero4BG and P-
Mero4SA, for sensing SNAP-tag and hCAII proteins. In the
presence of the target proteins, the probes showed dramatic
fluorescence increase of up to 300-fold for SNAP-tag and 70-
fold for hCAII. With P-Mero4BG, no-wash labeling of intra-
cellular proteins fused with SNAP-tag was successfully
achieved and the localization of the protein was specifically
visualized with minimal background staining. Furthermore, P-
Mero4BG and NAPH-Mero4SBG can also be harnessed to
image the incomplete Lamin A processing in living cells and
for in vivo imaging. We believe that these near-IR fluorescent-
switchable P-Mero4 dye derivatives can spearhead the design
and development of novel fluorogenic probes to become an
important tool in target-specific imaging in living cells and
animals.
S.; Richter, V.; Bruns, T.; Strauss, W. S.; Wittig, R. J. Microsc. 2012,
245, 311.
(8)
(9)
685.
(10) Umezawa, K.; Citterio, D.; Suzuki, K. Anal. Sci. 2014, 30,
327.
(11) Yasueda, Y.; Tamura, T.; Fujisawa, A.; Kuwata, K.; Tsuki-
ji, S.; Kiyonaka, S.; Hamachi, I. J. Am. Chem. Soc. 2016, 138, 7592.
(12) Zhu, H.; Fan, J.; Du, J.; Peng, X. Acc. Chem. Res. 2016, 49,
2115.
(13) Liu, Y.; Zhou, J.; Wang, L.; Hu, X.; Liu, X.; Liu, M.; Cao,
Z.; Shangguan, D.; Tan, W. J. Am. Chem. Soc. 2016, 138, 12368.
(14) Kim, E.; Yang, K. S.; Kohler, R. H.; Dubach, J. M.; Miku-
la, H.; Weissleder, R. Bioconjugate Chem. 2015, 26, 1513.
(15) Wombacher, R.; Heidbreder, M.; van de Linde, S.; Sheetz,
M. P.; Heilemann, M.; Cornish, V. W.; Sauer, M. Nat. Methods 2010,
7, 717.
(16) van de Linde, S.; Heilemann, M.; Sauer, M. Annu. Rev.
Phys. Chem. 2012, 63, 519.
(17) Loving, G. S.; Sainlos, M.; Imperiali, B. Trends Biotech-
nol. 2010, 28, 73.
1
2
3
4
5
6
7
8
Lavis, L. D.; Raines, R. T. ACS Chem. Biol. 2014, 9, 855.
Kushida, Y.; Nagano, T.; Hanaoka, K. Analyst 2015, 140,
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
(18) Klymchenko, A. S. Acc. Chem. Res. 2017, 50, 366.
(19) Toutchkine, A.; Kraynov, V.; Hahn, K. J. Am. Chem. Soc.
2003, 125, 4132.
(20) Fink, A. L.; Calciano, L. J.; Goto, Y.; Kurotsu, T.; Palleros,
D. R. Biochemistry 1994, 33, 12504.
(21) de Lorimier, R. M.; Smith, J. J.; Dwyer, M. A.; Looger, L.
L.; Sali, K. M.; Paavola, C. D.; Rizk, S. S.; Sadigov, S.; Conrad, D.
W.; Loew, L.; Hellinga, H. W. Protein Sci. 2002, 11, 2655.
(22) Nalbant, P.; Hodgson, L.; Kraynov, V.; Toutchkine, A.;
Hahn, K. M. Science 2004, 305, 1615.
(23) Sainlos, M.; Iskenderian, W. S.; Imperiali, B. J. Am. Chem.
Soc. 2009, 131, 6680.
(24) Zhuang, Y.-D.; Chiang, P.-Y.; Wang, C.-W.; Tan, K.-T.
Angew. Chem. Int. Ed. 2013, 52, 8124.
(25) Liu, T.-K.; Hsieh, P.-Y.; Zhuang, Y.-D.; Hsia, C.-Y.;
Huang, C.-L.; Lai, H.-P.; Lin, H.-S.; Chen, I. C.; Hsu, H.-Y.; Tan, K.-
T. ACS Chem. Biol. 2014, 9, 2359.
(26) Unger-Angel, L.; Rout, B.; Ilani, T.; Eisenstein, M.; Motiei,
L.; Margulies, D. Chem. Sci. 2015, 6, 5419.
(27) Baranczak, A.; Liu, Y.; Connelly, S.; Du, W.-G. H.;
Greiner, E. R.; Genereux, J. C.; Wiseman, R. L.; Eisele, Y. S.; Brad-
bury, N. C.; Dong, J.; Noodleman, L.; Sharpless, K. B.; Wilson, I. A.;
Encalada, S. E.; Kelly, J. W. J. Am. Chem. Soc. 2015, 137, 7404.
(28) Hori, Y.; Norinobu, T.; Sato, M.; Arita, K.; Shirakawa, M.;
Kikuchi, K. J. Am. Chem. Soc. 2013, 135, 12360.
(29) Chen, C.; Song, Z.; Zheng, X.; He, Z.; Liu, B.; Huang, X.;
Kong, D.; Ding, D.; Tang, B. Z. Chem. Sci. 2017, 8, 2191.
(30) Shirinian, V. Z.; Shimkin, A. A. In Heterocyclic Polyme-
thine Dyes: Synthesis, Properties and Applications; Strekowski, L.,
Ed.; Springer Berlin Heidelberg: Berlin, Heidelberg, 2008, p 75.
(31) Stillwell, W.; Wassall, S. R.; Dumaual, A. C.; Ehringer, W.
D.; Browning, C. W.; Jenski, L. J. Biochim. Biophys. Acta. 1993,
1146, 136.
(32) Hahn, K.; DeBiasio, R.; Taylor, D. L. Nature 1992, 359,
736.
(33) Woodward, R. B. J. Am. Chem. Soc. 1941, 63, 1123.
(34) Fieser, L. F.; Fieser, M.; Rajagopalan, S. J. Org. Chem.
1948, 13, 800.
(35) Pavia, D. L.; Lampman, G. M.; Kriz, G. S.; Vyvyan, J. A.
Introduction to Spectroscopy, 5th Edition; Cengage Learning: Boston,
2014.
(36) Krishnamurthy, V. M.; Kaufman, G. K.; Urbach, A. R.;
Gitlin, I.; Gudiksen, K. L.; Weibel, D. B.; Whitesides, G. M. Chem.
Rev. 2008, 108, 946.
ASSOCIATED CONTENT
Supporting Information. For complete experimental methods
and figures see Supplementary Information. This material is
AUTHOR INFORMATION
Corresponding Author
Notes
The authors declare no competing financial interests.
ACKNOWLEDGMENT
We are grateful to the Ministry of Science and Technology (Grant
No.: 106-2113-M-007-027) Taiwan (ROC) for financial support.
We thank Prof. Lily Hui-Ching Wang and Mr. Tian-Neng Li for
preparing the stable SNAP-PDGFR HeLa cells. Authors also
thank the Animal Molecular Imaging Core Facility and Miss Shu-
Hsien Wu for technical support of animal care in NHRI
(MOHW106-TDU-B-211-144-001 and NHRI-BN-106-PP-31).
We also thank Miss Li-Ching Shen of the Center for Advanced
Instrumentation of National Chiao Tung University for the 2D
NMR measurements.
REFERENCES
(1)
(2)
Waggoner, A. Curr. Opin. Chem. Biol. 2006, 10, 62.
Goddard, J. P.; Reymond, J. L. Curr. Opin. Biotechnol.
2004, 15, 314.
(3)
Kobayashi, H.; Ogawa, M.; Alford, R.; Choyke, P. L.; Ura-
no, Y. Chem. Rev. 2010, 110, 2620.
(4)
Spence, M. T. Z.; Johnson, I. D. The Molecular Probes
Handbook: A Guide to Fluorescent Probes and Labeling Technolo-
gies, 11th Edition; Life Technologies Corporation: New York, 2010.
(37) Keppler, A.; Gendreizig, S.; Gronemeyer, T.; Pick, H.; Vo-
gel, H.; Johnsson, K. Nat. Biotechnol. 2003, 21, 86.
(38) Kulinich, A. V.; Derevyanko, N. A.; Mikitenko, E. K.;
Ishchenko, A. A. J. Phys. Org. Chem. 2011, 24, 732.
(5)
(6)
Weissleder, R. Nat. Biotechnol. 2001, 19, 316.
Escobedo, J. O.; Rusin, O.; Lim, S.; Strongin, R. M. Curr.
Opin. Chem. Biol. 2010, 14, 64.
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