Monitoring autophagic degradation of p62/SQSTM1.
作者： Geir Bjørkøy , Trond Lamark , Serhiy Pankiv , Aud Øvervatn , Andreas Brech
DOI: 10.1016/S0076-6879(08)03612-4
关键词:
摘要: The p62 protein, also called sequestosome 1 (SQSTM1), is a ubiquitin-binding scaffold protein that colocalizes with ubiquitinated aggregates in many neurodegenerative diseases and proteinopathies of the liver. able to polymerize via an N-terminal PB1 domain can interact proteins C-terminal UBA domain. Also, p62/SQSTM1 binds directly LC3 GABARAP family specific sequence motif. itself degraded by autophagy may serve link autophagic machinery enable their degradation lysosome. Since accumulates when inhibited, decreased levels be observed induced, used as marker study flux. Here, we present several protocols for monitoring autophagy-mediated using Western blots, pulse-chase measurement half-life, immunofluorescence immuno-electron microscopy, well live cell imaging pH-sensitive mCherry-GFP double tag. We data on species-specificity map epitopes recognized commercially available anti-p62 antibodies.
sci-hub.se 参考文章(29)
Noboru Mizushima, Tamotsu Yoshimori, How to Interpret LC3 Immunoblotting Autophagy. ,vol. 3, pp. 542- 545 ,(2007) , 10.4161/AUTO.4600
Nigel Chaffey, Red fluorescent protein Trends in Plant Science. ,vol. 6, pp. 98- ,(2001) , 10.1016/S1360-1385(01)01902-1
E. Kuusisto, T. Kauppinen, I. Alafuzoff, Use of p62/SQSTM1 antibodies for neuropathological diagnosis Neuropathology and Applied Neurobiology. ,vol. 34, pp. 169- 180 ,(2008) , 10.1111/J.1365-2990.2007.00884.X
M. Lamar Seibenhener, Thangiah Geetha, Marie W. Wooten, Sequestosome 1/p62 – More than just a scaffold FEBS Letters. ,vol. 581, pp. 175- 179 ,(2007) , 10.1016/J.FEBSLET.2006.12.027
M. Olivé, F. W. van Leeuwen, A. Janué, D. Moreno, B. Torrejón-Escribano, I. Ferrer, Expression of mutant ubiquitin (UBB+1) and p62 in myotilinopathies and desminopathies. Neuropathology and Applied Neurobiology. ,vol. 34, pp. 76- 87 ,(2007) , 10.1111/J.1365-2990.2007.00864.X
Peter J. Peters, Jacques J. Neefjes, Viola Oorschot, Hidde L. Ploegh, Hans J. Geuze, Segregation of MHC class II molecules from MHC class I molecules in the Golgi complex for transport to lysosomal compartments. Nature. ,vol. 349, pp. 669- 676 ,(1991) , 10.1038/349669A0
Serhiy Pankiv, Terje Høyvarde Clausen, Trond Lamark, Andreas Brech, Jack-Ansgar Bruun, Heidi Outzen, Aud Øvervatn, Geir Bjørkøy, Terje Johansen, p62/SQSTM1 Binds Directly to Atg8/LC3 to Facilitate Degradation of Ubiquitinated Protein Aggregates by Autophagy Journal of Biological Chemistry. ,vol. 282, pp. 24131- 24145 ,(2007) , 10.1074/JBC.M702824200
Jozsef Gal, Anna-Lena Ström, Renee Kilty, Fujian Zhang, Haining Zhu, p62 accumulates and enhances aggregate formation in model systems of familial amyotrophic lateral sclerosis. Journal of Biological Chemistry. ,vol. 282, pp. 11068- 11077 ,(2007) , 10.1074/JBC.M608787200
Atsuko Nakai, Osamu Yamaguchi, Toshihiro Takeda, Yoshiharu Higuchi, Shungo Hikoso, Masayuki Taniike, Shigemiki Omiya, Isamu Mizote, Yasushi Matsumura, Michio Asahi, Kazuhiko Nishida, Masatsugu Hori, Noboru Mizushima, Kinya Otsu, The role of autophagy in cardiomyocytes in the basal state and in response to hemodynamic stress. Nature Medicine. ,vol. 13, pp. 619- 624 ,(2007) , 10.1038/NM1574
Erkki Kuusisto, Antero Salminen, Irina Alafuzoff, Ubiquitin-binding protein p62 is present in neuronal and glial inclusions in human tauopathies and synucleinopathies. Neuroreport. ,vol. 12, pp. 2085- 2090 ,(2001) , 10.1097/00001756-200107200-00009