Monitoring β-Secretase Activity in Living Cells with a Membrane-Anchored FRET Probe†
Drew S. Folk
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorJustin C. Torosian
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorSunhee Hwang
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorProf. Dr. Dewey G. McCafferty
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorCorresponding Author
Prof. Katherine J. Franz
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)Search for more papers by this authorDrew S. Folk
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorJustin C. Torosian
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorSunhee Hwang
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorProf. Dr. Dewey G. McCafferty
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Search for more papers by this authorCorresponding Author
Prof. Katherine J. Franz
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)
Department of Chemistry, Duke University, 124 Science Dr. Durham, NC 27708 (USA)Search for more papers by this authorWe thank the National Institutes of Health (grant GM084176), the Sloan Foundation, and the Camille and Henry Dreyfus Foundation for supporting this work. Special thanks to Sam Johnson and the Duke LMCF (grant 1S10RR027528-01) for their microscopy expertise.
Graphical Abstract
Turn it on! β-MAP is a sensitive FRET probe with specificity for monitoring the enzyme β-secretase (BACE), which is associated with Alzheimer's disease. After hydrolysis by the enzyme BACE, the probe fluoresces and thus allows real-time spatial and temporal assessment of enzymatic activity in living cells. β-MAP was used to confirm the cellular efficacy of a reported inhibitor without the need for mutated cell lines or antibodies.
Supporting Information
Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors.
| Filename | Description |
|---|---|
| anie_201206673_sm_miscellaneous_information.pdf1.7 MB | miscellaneous_information |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
References
- 1J. Hardy, D. Selkoe, Science 2002, 297, 353.
- 2
- 2aA. Rauk, Dalton Trans. 2008, 1273;
- 2bP. Crouch, S. Harding, A. White, J. Camakaris, A. Bush, C. Masters, Int. J. Biochem. Cell Biol. 2008, 40, 181.
- 3G. Evin, A. Barakat, C. Masters, Int. J. Biochem. Cell Biol. 2010, 42, 1923.
- 4V. Frisardi et al., Curr. Alzheimer Res. 2010, 7, 40. See the Supporting Information for all authors.
- 5R. Postina, Curr. Alzheimer Res. 2008, 5, 179.
- 6K. Endres, F. Fahrenholz, FEBS J. 2010, 277, 1585.
- 7M. Hutton, J. Perez-Tur, J. Hardy, Essays Biochem. 1998, 33, 117.
- 8
- 8aM. Hemming, J. Elias, S. Gygi, D. Selkoe, PLoS One 2009, 4;
- 8bF. Gruninger-Leitch, D. Schlatter, E. Kung, P. Nelbock, H. Dobeli, J. Biol. Chem. 2002, 277, 4687.
- 9F. Mancini, A. De Simone, V. Andrisano, Anal. Bioanal. Chem. 2011, 400, 1979.
- 10
- 10aM. Congreve et al., J. Med. Chem. 2007, 50, 1124;
- 10bY. Cheng et al., J. Med. Chem. 2011, 54, 5836;
- 10cD. Huang, U. Luthi, P. Kolb, M. Cecchini, A. Barberis, A. Caflisch, J. Am. Chem. Soc. 2006, 128, 5436;
- 10dY. Shimmyo, T. Kihara, A. Akaike, T. Niidome, H. Sugimoto, Biochim. Biophys. Acta Gen. Subj. 2008, 1780, 819;
- 10eW. Yang et al., J. Med. Chem. 2006, 49, 839. See the Supporting Information for all authors.
- 11
- 11aS. B. Halima, L. Rajendran, J. Alzheimer′s Dis. 2011, 24, 143;
- 11bS. Stachel et al., Bioorg. Med. Chem. Lett. 2009, 19, 2977. See the Supporting Information for all authors.
- 12
- 12aS. Stachel et al., J. Med. Chem. 2004, 47, 6447;
- 12bX. P. Shi et al., J. Alzheimer′s Dis. 2005, 7, 139;
- 12cH. Yamakawa, S. Yagishita, E. Futai, S. Ishiura, J. Biol. Chem. 2010, 285, 1634;
- 12dM. Oh, S. Kim, Y. Oh, D. Choi, H. Sin, I. Jung, W. Park, Anal. Biochem. 2003, 323, 7;
- 12eB. Pietrak et al., Anal. Biochem. 2005, 342, 144. See the Supporting Information for all authors.
- 13J. Lu, Z. Zhang, J. Yang, J. Chu, P. Li, S. Zeng, Q. Luo, Biochem. Biophys. Res. Commun. 2007, 362, 25.
- 14
- 14aA. Cobos-Correa, J. B. Trojanek, S. Diemer, M. A. Mall, C. Schultz, Nat. Chem. Biol. 2009, 5, 628;
- 14bE. A. Lemke, C. Schultz, Nat. Chem. Biol. 2011, 7, 480.
- 15L. Rajendran et al., Science 2008, 320, 520. See the Supporting Information for all authors.
- 16D. S. Folk, K. J. Franz, J. Am. Chem. Soc. 2010, 132, 4994.
- 17A. Laude, I. Prior, Mol. Membr. Biol. 2004, 21, 193.
- 18
- 18aS. Kim, J. Yi, Y. Ko, J. Cell. Biochem. 2006, 99, 878;
- 18bD. Riddell, G. Christie, I. Hussain, C. Dingwall, Curr. Biol. 2001, 11, 1288.
- 19J. Lenhart, X. Ling, R. Gandhi, T. Guo, P. Gerk, D. Burnzell, S. Zhang, J. Med. Chem. 2010, 53, 6198.
- 20
- 20aD. Andrau et al., J. Biol. Chem. 2003, 278, 25859;
- 20bJ. Ermolieff, J. Loy, G. Koelsch, J. Tang, Biochemistry 2000, 39, 12450. See the Supporting Information for all authors of Ref. [20 a].
- 21K. C. Wilcox, P. N. Lacor, J. Pitt, W. L. Klein, Cell. Mol. Neurobiol. 2011, 31, 939.
