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The Carboxyl-Terminal Amino Acids Render Pro-Human LC3B Migration Similar to Lipidated LC3B in SDS-PAGE

Wang, W and Chen, Z and Billiar, TR and Stang, MT and Gao, W (2013) The Carboxyl-Terminal Amino Acids Render Pro-Human LC3B Migration Similar to Lipidated LC3B in SDS-PAGE. PLoS ONE, 8 (9).

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Abstract

LC3 is widely used marker for macroautophagy assays. After translation pro-LC3 is processed by Atg4 to expose C-terminal glycine residue for downstream conjugation reactions to accomplish the conversion of LC3-I to LC3-II. SDS-PAGE based Western blot (Wb) is generally utilized to quantify LC3-II levels where the LC3-I band migrates slower than LC3-II. We found that pro-human LC3B migrated at similar rate as LC3B-II in SDS-PAGE. The carboxyl-terminal five amino acids, particularly Lysine122 and Leucine123 of human LC3B play a major role in the faster migration of unprocessed LC3B, rendering it indistinguishable from LC3B-II in Wb assays. The unique faster migration of unprocessed LC3B than LC3B-I is also revealed in mouse LC3B, rat LC3B and rat LC3 but not in human LC3C. Our findings for the first time define pro-LC3 migration patterns for LC3 family member from human, mouse and rat species in SDS-PAGE. These findings provide a reference for pro-LC3 band patterns when Atg4 function is inhibited. © 2013 Wang et al.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Wang, W
Chen, Z
Billiar, TRbilliar@pitt.eduBILLIAR
Stang, MT
Gao, Wgaowent@pitt.eduGAOWENT
Date: 10 September 2013
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 8
Number: 9
DOI or Unique Handle: 10.1371/journal.pone.0074222
Schools and Programs: School of Medicine > Surgery
Refereed: Yes
Date Deposited: 11 Oct 2013 19:04
Last Modified: 19 Feb 2019 15:55
URI: http://d-scholarship.pitt.edu/id/eprint/19832

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