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Use of Peptide Arrays for Identification and Characterization of LIR Motifs

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https://hdl.handle.net/10037/17721
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Date
2019-01-05
Type
Journal article
Tidsskriftartikkel
Peer reviewed

Author
Rasmussen, Mads Skytte; Birgisdottir, Åsa birna; Johansen, Terje
Abstract
The mammalian ATG8 proteins (LC3A-C/GABARAP, GABARAPL1, and GABARAPL2) are small ubiquitin-like proteins critically involved in macroautophagy. Their processed C-termini are posttranslationally conjugated to a phosphatidylethanolamine moiety, enabling their insertion into the lipid bilayers of both the inner and outer membranes of the forming autophagosomes. The ATG8s bind a diverse selection of proteins including cargo receptors for selective autophagy, members of the core autophagy machinery, and other proteins involved in formation, transport, and maturation (fusion to lysosomes) of autophagosomes. Protein binding to the ATG8s is in most cases mediated by short, conserved sequence motifs known as LC3-interacting regions (LIRs). Here, we present a protocol for identifying putative LIR motifs in a whole protein sequence using peptide arrays generated by SPOT synthesis on nitrocellulose membranes. The use of two-dimensional peptide arrays allows for further identification of specific residues critical for LIR binding.
Description
This is a post-peer-review, pre-copyedit version of an article published in Methods in Molecular Biology. The final authenticated version is available online at: http://dx.doi.org/10.1007/978-1-4939-8873-0_8
Publisher
Springer Nature
Citation
Rasmussen, M.S., Birgisdottir, Å.b., Johansen, T. (2019) Use of Peptide Arrays for Identification and Characterization of LIR Motifs. Methods in molecular biology, 1880, 149-161
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© Springer Science+Business Media, LLC, part of Springer Nature 2019

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