Atg16L1 N-terminal Antibody (Rabbit mAb) [E9G16] Datasheet

Biological Description

Specificity	Atg16L1 N-terminal Antibody (Rabbit mAb) [E9G16] detects endogenous levels of total Atg16L1 protein.
Background	ATG16L1 (autophagy-related 16-like 1) is the mammalian counterpart of yeast Atg16 and an essential component of the machinery that builds autophagosomes. It is expressed in all tissues and pairs a short N-terminal region with a coiled-coil domain, followed by a C-terminal WD-repeat domain that yeast Atg16 lacks. The N-terminal portion carries its core functions. The extreme N-terminal segment, just upstream of the coiled-coil, is enough to bind ATG5. ATG16L1 binds ATG5 whether or not ATG5 is conjugated to ATG12, which points to direct contact with ATG5 rather than ATG12. The N-terminal region including the coiled-coil drives self-association, and full-length ATG16L1 oligomerizes independently of ATG5. The WD-repeat domain is dispensable for both ATG5 binding and oligomerization. Through these N-terminal interactions, ATG16L1 and the ATG12–ATG5 conjugate assemble into a large multimeric complex. Without ATG5, this complex does not form, and ATG16L1 stays as a smaller, presumably tetrameric unit. Most ATG16L1 is in the cytosol. During amino acid starvation it forms puncta and moves with ATG5 to small vesicles and early isolation membranes, collecting mainly on the outer face of the cup-shaped isolation membrane. It leaves once the autophagosome is complete and is absent from LC3-positive mature autophagosomes and autolysosomes. Its membrane targeting needs ATG5 binding but not ATG12 conjugation. The ATG16L1 complex acts as an E3-like scaffold for LC3 lipidation. The site where LC3 is conjugated to phosphatidylethanolamine depends on where the ATG16L1 complex sits on membranes and on the interaction of ATG12 with ATG3, the E2 enzyme for LC3. Forcing ATG16L1 to the plasma membrane moves LC3 lipidation there, and overexpressing ATG16L1 or ATG12 inhibits autophagosome formation. Alternative splicing of exons between the coiled-coil and WD-repeat regions produces α, β and γ isoforms. The α form predominates in liver and the γ form in brain, and all three join the large complex.

Specificity

Atg16L1 N-terminal Antibody (Rabbit mAb) [E9G16] detects endogenous levels of total Atg16L1 protein.

Background

ATG16L1 (autophagy-related 16-like 1) is the mammalian counterpart of yeast Atg16 and an essential component of the machinery that builds autophagosomes. It is expressed in all tissues and pairs a short N-terminal region with a coiled-coil domain, followed by a C-terminal WD-repeat domain that yeast Atg16 lacks. The N-terminal portion carries its core functions. The extreme N-terminal segment, just upstream of the coiled-coil, is enough to bind ATG5. ATG16L1 binds ATG5 whether or not ATG5 is conjugated to ATG12, which points to direct contact with ATG5 rather than ATG12. The N-terminal region including the coiled-coil drives self-association, and full-length ATG16L1 oligomerizes independently of ATG5. The WD-repeat domain is dispensable for both ATG5 binding and oligomerization. Through these N-terminal interactions, ATG16L1 and the ATG12–ATG5 conjugate assemble into a large multimeric complex. Without ATG5, this complex does not form, and ATG16L1 stays as a smaller, presumably tetrameric unit. Most ATG16L1 is in the cytosol. During amino acid starvation it forms puncta and moves with ATG5 to small vesicles and early isolation membranes, collecting mainly on the outer face of the cup-shaped isolation membrane. It leaves once the autophagosome is complete and is absent from LC3-positive mature autophagosomes and autolysosomes. Its membrane targeting needs ATG5 binding but not ATG12 conjugation. The ATG16L1 complex acts as an E3-like scaffold for LC3 lipidation. The site where LC3 is conjugated to phosphatidylethanolamine depends on where the ATG16L1 complex sits on membranes and on the interaction of ATG12 with ATG3, the E2 enzyme for LC3. Forcing ATG16L1 to the plasma membrane moves LC3 lipidation there, and overexpressing ATG16L1 or ATG12 inhibits autophagosome formation. Alternative splicing of exons between the coiled-coil and WD-repeat regions produces α, β and γ isoforms. The α form predominates in liver and the γ form in brain, and all three join the large complex.

Usage Information

Application

WB, IF

Dilution

WB	IF
1:1000	1:50-1:100

Reactivity

Mouse, Rat, Human

Source

Rabbit Monoclonal Antibody

MW

68 kDa

Storage Buffer

PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage
(from the date of receipt)

-20°C (avoid freeze-thaw cycles), 2 years

References

https://pubmed.ncbi.nlm.nih.gov/12665549/
https://pubmed.ncbi.nlm.nih.gov/18321988/

Atg16L1 N-terminal Antibody (Rabbit mAb) [E9G16]

Biological Description

Usage Information

References

Application Data