What is LC3 in autophagy?
What is LC3 in autophagy?
LC3 is a central protein in the autophagy pathway where it functions in autophagy substrate selection and autophagosome biogenesis. LC3 is the most widely used marker of autophagosomes.
What does it mean when LC3 Atg8 becomes Lipidated?
Atg8/LC3membrane association is mediated by the covalent attachment of Atg8/LC3 to phosphatidylethanolamine (PE)2, 3. The resulting lipidated protein then drives autophagosome maturation steps including cargo capture, growth and closure of the organelle4, 5, and lysosome targeting or recognition (for review6).
How do you calculate autophagy?
One of the most common ways to monitor autophagy is by measuring the protein levels of LC3-II, which is incorporated into autophagosomes and then degraded in the lysosome. There are many different ways to measure LC3 protein levels, including western blot and immunofluorescent microscopy2.
Can you measure autophagy?
Autophagy can be monitored by two different approaches: (1) direct observation of autophagy-related structures and their fate; and (2) quantification of autophagy-/lysosome-dependent degradation of proteins and organelles.
What happens to LC3 during autophagy?
During autophagy, autophagosomes engulf cytoplasmic components, including cytosolic proteins and organelles. Autophagosomes fuse with lysosomes to form autolysosomes, and intra-autophagosomal components are degraded by lysosomal hydrolases. At the same time, LC3-II in autolysosomal lumen is degraded.
Is ATG8 same as LC3?
ATG8/LC3 family: expression & structure. Yeast Atg8, is an ubiquitin-like (Ubl) protein, exhibiting amino acid sequence similarity with LC3 (A, B, C), GABARAP, GABARAPL1(GEC1), GABARAPL2 (also known as GATE-16 (Golgi-associated ATPase enhancer of 16 kDa)), and GABARAPL3 (Fig.
What is the known function of ATG8?
For example, ATG8 proteins regulate membrane elongation during autophagosome biogenesis. Moreover, they play a crucial role in cargo recognition for selective autophagy by interacting with autophagy receptors/adaptors. In addition to their autophagic roles, ATG8 proteins play non-autophagic roles in plants.
How do you quantify LC3?
To quantify it, measure the flux by detecting the difference in LC3-II levels in the presence vs the absence of lysosomal inhibitors. For cell cultures, don’t forget your controls, because you will need two wells for each treatment, one with and one without inhibitors.
How many autophagosomes are in a cell?
Table 1.
| Variable | Unit | Induced |
|---|---|---|
| Autophagosome flux, J | A/h/cell | 105.4 |
| Number of autophagosomes, nA | A/cell | 16.7 |
| Number of autolysosomes, nAL | AL/cell | 251.2 |
| Average volume of an autophagosome | fL | 4.7 |
Why are LC3/GABARAP proteins important in autophagy?
Giventhe high homology, the other LC3/GABARAP family proteins are often presumed to fulfill similar functions. Nevertheless, substantial evidence shows that the LC3/GABARAP family proteins are unique in function and important in autophagy-independent mechanisms.
What is the difference between LC3A-C and GABARAP?
LC3A-C (LC3A has two variants originating from alternative splicing event) belong to the LC3 subfamily while GABARAP, GABARAPL1, GABARAPL2 (GATE-16), and GABARAPL3 are members of the GABARAP/GATE-16 subfamily. LC3 and GABARAP/GATE-16 subfamilies are both essential in autophagosome biogenesis but have different functions.
What is the role of GABARAPL2 in Uba5?
An atypical LIR motif within UBA5 (ubiquitin like modifier activating enzyme 5) interacts with GABARAP proteins and mediates membrane localization of UBA5. GABARAPL2 Is Critical for Growth Restriction of Toxoplasma gondii in HeLa Cells Treated with Gamma Interferon.
Can ravz protein interfere with autophagy in Legionella pneumophila?
ATG8 proteins act as scaffolds for assembly of the ULK complex at the phagophore Legionella pneumophila could interfere with autophagy by using the bacterial effector protein RavZ to directly uncouple Atg8 proteins attached to phosphatidylethanolamine on autophagosome membranes.
