Skip to Content
Merck

Phosphorylated ubiquitin chain is the genuine Parkin receptor.

The Journal of cell biology (2015-04-08)
Kei Okatsu, Fumika Koyano, Mayumi Kimura, Hidetaka Kosako, Yasushi Saeki, Keiji Tanaka, Noriyuki Matsuda
ABSTRACT

PINK1 selectively recruits Parkin to depolarized mitochondria for quarantine and removal of damaged mitochondria via ubiquitylation. Dysfunction of this process predisposes development of familial recessive Parkinson's disease. Although various models for the recruitment process have been proposed, none of them adequately explain the accumulated data, and thus the molecular basis for PINK1 recruitment of Parkin remains to be fully elucidated. In this study, we show that a linear ubiquitin chain of phosphomimetic tetra-ubiquitin(S65D) recruits Parkin to energized mitochondria in the absence of PINK1, whereas a wild-type tetra-ubiquitin chain does not. Under more physiologically relevant conditions, a lysosomal phosphorylated polyubiquitin chain recruited phosphomimetic Parkin to the lysosome. A cellular ubiquitin replacement system confirmed that ubiquitin phosphorylation is indeed essential for Parkin translocation. Furthermore, physical interactions between phosphomimetic Parkin and phosphorylated polyubiquitin chain were detected by immunoprecipitation from cells and in vitro reconstitution using recombinant proteins. We thus propose that the phosphorylated ubiquitin chain functions as the genuine Parkin receptor for recruitment to depolarized mitochondria.

MATERIALS
Product Number
Brand
Product Description

Supelco
Sodium chloride, Pharmaceutical Secondary Standard; Certified Reference Material
Sorbitol, European Pharmacopoeia (EP) Reference Standard
USP
Sorbitol, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Acrylamide, for molecular biology, ≥99% (HPLC)
Sigma-Aldrich
Sodium chloride solution, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Sodium pyruvate, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99%
Sigma-Aldrich
Sodium pyruvate, BioXtra, ≥99%
Sigma-Aldrich
Sodium pyruvate, powder, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
Sodium pyruvate, Hybri-Max, powder, suitable for hybridoma
Sigma-Aldrich
Sodium pyruvate, ReagentPlus®, ≥99%
Sigma-Aldrich
Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Sodium chloride, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Sodium chloride, tablet
Sigma-Aldrich
Sodium chloride, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
Monoclonal Anti-Actin antibody produced in mouse, clone AC-40, ascites fluid
Sigma-Aldrich
D-Sorbitol, ≥98% (GC), for molecular biology
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Tetracycline, 98.0-102.0% (HPLC)
Sigma-Aldrich
D-Sorbitol, ≥98% (GC), BioReagent, suitable for cell culture, suitable for plant cell culture
Sigma-Aldrich
Sodium chloride solution, 5 M
Sigma-Aldrich
D-Sorbitol, ≥98% (GC), BioXtra
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99% (HPLC), powder
Sigma-Aldrich
Tetracycline, 98.0-102.0% (HPLC)
Sigma-Aldrich
Sodium chloride solution, BioUltra, for molecular biology, ~5 M in H2O
Sigma-Aldrich
D-Sorbitol, BioUltra, ≥99.0% (HPLC)
Sigma-Aldrich
Sodium chloride, tested according to Ph. Eur.
Supelco
Sodium chloride, reference material for titrimetry, certified by BAM, >99.5%
Sigma-Aldrich
Acrylamide, purum, ≥98.0% (GC)
Sigma-Aldrich
Sodium chloride, AnhydroBeads, −10 mesh, 99.999% trace metals basis
Sigma-Aldrich
Sodium chloride, random crystals, optical grade, 99.9% trace metals basis