Document Detail

Cytosolic LC3 ratio as a quantitative index of macroautophagy.
MedLine Citation:
PMID:  19200884     Owner:  NLM     Status:  MEDLINE    
Macroautophagy, an intracellular bulk degradation process and a typical form of autophagy in eukaryotes, is sensitive to physiological regulation, such as the supply and deprivation of nutrients. Microtubule-associated protein 1 light chain 3 (LC3), a mammalian homologue of yeast Atg8, plays a critical role in macroautophagy formation and is considered a suitable marker for this process. In mammalian cells, there is a limitation for biochemical and morphological methods to monitor autophagy within a short period of time. During analysis of the subcellular distribution of LC3, we found that the cytosolic fraction contains not only a precursor form (LC3-I), but also an apparently active form, denoted as LC3-IIs. Both LC3-I and LC3-IIs in the cytosolic fraction, and thus the LC3-IIs/I ratio (designated the cytosolic LC3 ratio), were more responsive to amino acids than monitoring LC3-II or the LC3-II/I ratio in the total homogenate, and remarkably reflected the total proteolytic flux in fresh rat hepatocytes and the cultured H4-II-E cell line. Thus, in addition to representing a sensitive index of macroautophagy, examining the cytosolic LC3 ratio is an easy and quick quantitative method for monitoring the regulation of this process in hepatocytes and H4-II-E cells.
Motoni Kadowaki; Md Razaul Karim
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in enzymology     Volume:  452     ISSN:  1557-7988     ISO Abbreviation:  Meth. Enzymol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-02-09     Completed Date:  2009-04-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0212271     Medline TA:  Methods Enzymol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  199-213     Citation Subset:  IM    
Department of Applied Biological Chemistry, Faculty of Agriculture, Graduate School of Science and Technology, Niigata University, Niigata, Japan.
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MeSH Terms
Autophagy / physiology*
Cell Line
Cells, Cultured
Cytosol / metabolism*
Hepatocytes / metabolism
Microtubule-Associated Proteins / physiology*
Reg. No./Substance:
0/LC3 protein, rat; 0/Microtubule-Associated Proteins

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