Ub-ProT reveals global length and composition of protein ubiquitylation in cells

Hikaru Tsuchiya, Daocharad Burana, Fumiaki Ohtake, Naoko Arai, Ai Kaiho, Masayuki Komada, Keiji Tanaka () and Yasushi Saeki ()
Additional contact information
Hikaru Tsuchiya: Tokyo Metropolitan Institute of Medical Science
Daocharad Burana: Tokyo Metropolitan Institute of Medical Science
Fumiaki Ohtake: Tokyo Metropolitan Institute of Medical Science
Naoko Arai: Tokyo Metropolitan Institute of Medical Science
Ai Kaiho: Tokyo Metropolitan Institute of Medical Science
Masayuki Komada: Tokyo Institute of Technology
Keiji Tanaka: Tokyo Metropolitan Institute of Medical Science
Yasushi Saeki: Tokyo Metropolitan Institute of Medical Science

Nature Communications, 2018, vol. 9, issue 1, 1-10

Abstract: Abstract Protein ubiquitylation regulates diverse cellular processes via distinct ubiquitin chains that differ by linkage type and length. However, a comprehensive method for measuring these properties has not been developed. Here we describe a method for assessing the length of substrate-attached polyubiquitin chains, “ubiquitin chain protection from trypsinization (Ub-ProT).” Using Ub-ProT, we found that most ubiquitylated substrates in yeast-soluble lysate are attached to chains of up to seven ubiquitin molecules. Inactivation of the ubiquitin-selective chaperone Cdc48 caused a dramatic increase in chain lengths on substrate proteins, suggesting that Cdc48 complex terminates chain elongation by substrate extraction. In mammalian cells, we found that ligand-activated epidermal growth factor receptor (EGFR) is rapidly modified with K63-linked tetra- to hexa-ubiquitin chains following EGF treatment in human cells. Thus, the Ub-ProT method can contribute to our understanding of mechanisms regulating physiological ubiquitin chain lengths and composition.

Date: 2018
References: Add references at CitEc
Citations: View citations in EconPapers (3)

Downloads: (external link)
https://www.nature.com/articles/s41467-018-02869-x Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-02869-x

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-018-02869-x

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().