6STF

Human Rab8a phosphorylated at Ser111 in complex with GDP

6STF の概要

• エントリーDOI: 10.2210/pdb6stf/pdb
• 関連するPDBエントリー: 4LHV
• 分子名称: Ras-related protein Rab-8A, GUANOSINE-5'-DIPHOSPHATE, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: parkinson disease, phosphorylation, small g-protein, sf3 motif, signaling protein
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 5
• 化学式量合計: 105966.02

構造登録者

Vieweg, S.,Mulholland, K.,Braeuning, B.,Kachariya, N.,Lai, Y.,Toth, R.,Sattler, M.,Groll, M.,Itzen, A.,Muqit, M.M.K. (登録日: 2019-09-10, 公開日: 2020-04-22, 最終更新日: 2024-10-16)

主引用文献

Vieweg, S.,Mulholland, K.,Brauning, B.,Kachariya, N.,Lai, Y.C.,Toth, R.,Singh, P.K.,Volpi, I.,Sattler, M.,Groll, M.,Itzen, A.,Muqit, M.M.K.
PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72.
Biochem.J., 477:1651-1668, 2020

PubMed Abstract: Loss of function mutations in the PTEN-induced kinase 1 (PINK1) kinase are causal for autosomal recessive Parkinson's disease (PD) whilst gain of function mutations in the LRRK2 kinase cause autosomal dominant PD. PINK1 indirectly regulates the phosphorylation of a subset of Rab GTPases at a conserved Serine111 (Ser111) residue within the SF3 motif. Using genetic code expansion technologies, we have produced stoichiometric Ser111-phosphorylated Rab8A revealing impaired interactions with its cognate guanine nucleotide exchange factor and GTPase activating protein. In a screen for Rab8A kinases we identify TAK1 and MST3 kinases that can efficiently phosphorylate the Switch II residue Threonine72 (Thr72) in a similar manner as LRRK2 in vitro. Strikingly, we demonstrate that Ser111 phosphorylation negatively regulates the ability of LRRK2 but not MST3 or TAK1 to phosphorylate Thr72 of recombinant nucleotide-bound Rab8A in vitro and demonstrate an interplay of PINK1- and LRRK2-mediated phosphorylation of Rab8A in transfected HEK293 cells. Finally, we present the crystal structure of Ser111-phosphorylated Rab8A and nuclear magnetic resonance structure of Ser111-phosphorylated Rab1B. The structures reveal that the phosphorylated SF3 motif does not induce any major changes, but may interfere with effector-Switch II interactions through intramolecular H-bond formation and/or charge effects with Arg79. Overall, we demonstrate antagonistic regulation between PINK1-dependent Ser111 phosphorylation and LRRK2-mediated Thr72 phosphorylation of Rab8A indicating a potential cross-talk between PINK1-regulated mitochondrial homeostasis and LRRK2 signalling that requires further investigation in vivo.

PubMed: 32227113
DOI: 10.1042/BCJ20190664

実験手法

X-RAY DIFFRACTION (2.4 Å)