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Cited by 56

The DOI matches its Crossref record; 6 of the 6 references it cites were found in a registry or an index.

Development of a novel method for quantification of autophagic protein degradation by AHA labeling

Zhang, Jianbin, Wang, Jigang, Ng, Shukie, Lin, Qingsong, Shen, Han-Ming·2014·Autophagy
DOI 10.4161/auto.28267
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Abstract

Autophagy is a catabolic process during which cellular components including protein aggregates and organelles are degraded via a lysosome-dependent process to sustain metabolic homeostasis during nutrient or energy deprivation. Measuring the rate of proteolysis of long-lived proteins is a classical assay for measurement of autophagic flux. However, traditional methods, such as a radioisotope labeling assay, are technically tedious and have low sensitivity. Here, we report a novel method for quantification of long-lived protein degradation based on L-azidohomoalanine (AHA) labeling in mouse embryonic fibroblasts (MEFs) and in human cancer cells. AHA is a surrogate for L-methionine, containing a bio-orthogonalazide moiety. When added to cultured cells, AHA is incorporated into proteins during active protein synthesis. After a click reaction between an azide and an alkyne, the azide-containing proteins can be detected with an alkyne-tagged fluorescent dye, coupled with flow cytometry. Induction of autophagy by starvation or mechanistic target of rapamycin (MTOR) inhibitors was able to induce a significant reduction of the fluorescence intensity, consistent with other autophagic markers. Coincidently, inhibition of autophagy by pharmacological agents or by Atg gene deletion abolished the reduction of the fluorescence intensity. Compared with the classical radioisotope pulse-labeling method, we think that our method is sensitive, quantitative, nonradioactive, and easy to perform,…

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  1. 10.1016/j.cell.2011.10.026 · DOI
  2. 10.1016/j.cell.2007.12.018 · DOI · Paper page
  3. 10.1152/ajpcell.00507.2009 · DOI
  4. 10.1101/gad.1599207 · DOI
  5. 10.1038/nrm2708 · DOI
  6. 10.4161/auto.8709 · DOI · Paper page
  7. 10.1016/j.cell.2012.03.017 · DOI · Paper page
  8. 10.1016/j.febslet.2010.01.017 · DOI · Paper page
  9. 10.1074/jbc.271.45.28593 · DOI
  10. 10.1007/978-1-59745-157-4_6 · DOI
  11. 10.1016/S0076-6879(08)03604-5 · DOI
  12. 10.1021/bi9007726 · DOI
  13. 10.1073/pnas.0601637103 · DOI · Paper page
  14. 10.1038/nprot.2007.52 · DOI
  15. 10.1073/pnas.012583299 · DOI
  16. 10.4161/auto.5662 · DOI
  17. 10.1016/j.bbrc.2006.09.168 · DOI
  18. 10.1038/cr.2013.11 · DOI
  19. 10.1074/jbc.275.2.992 · DOI
  20. 10.1126/science.1121925 · DOI
  21. 10.1038/nn.2580 · DOI
  22. 10.1021/cn2000876 · DOI
  23. 10.1042/CBI20110055 · DOI
  24. 10.1038/417177a · DOI
  25. 10.4161/auto.5939 · DOI
  26. 10.1016/j.febslet.2006.04.008 · DOI · Paper page
  27. 10.1038/nature03029 · DOI · Paper page
  28. 10.1083/jcb.200412022 · DOI · Paper page
  29. 10.1021/ja036765z · DOI
  30. 10.4161/auto.7.10.16681 · DOI · Paper page
  31. 10.1038/cdd.2010.72 · DOI

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Metadata

Authors
Zhang, J., Wang, J., Ng, S., Lin, Q., & Shen, H.-M.
Venue
Autophagy
Publisher
Informa UK Limited
Published
2014-03-24
Volume
10
Issue
5
Pages
901-912
ISSN
  • 1554-8627
  • 1554-8635
DOI
10.4161/auto.28267
Cited by
56
References
6
Language
en

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