PDBsum entry 5kty

Hydrolase

PDB id: 5kty

Contents

Protein chain

397 a.a.

Ligands

GDP ×2

Metals

_CA ×2

_MG

Waters ×76

PDB id:

5kty

Name:

Hydrolase

Title:

Hmiro ef hand and cgtpase domains, gdp and ca2+ bound state

Structure:

Mitochondrial rho gtpase 1. Chain: a. Fragment: hand and cgtpase domains (unp residues 177-592). Synonym: hmiro-1, rac-gtp-binding protein-like protein, ras homolog gene family member t1. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: rhot1, arht1. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.52Å R-factor: 0.213 R-free: 0.249

Authors:

J.L.Klosowiak,P.J.Focia,S.E.Rice,D.M.Freymann

Key ref:

J.L.Klosowiak et al. (2016). Structural insights into Parkin substrate lysine targeting from minimal Miro substrates. Sci Rep, 6, 33019. PubMed id: 27605430 DOI: 10.1038/srep33019

Date:

12-Jul-16 Release date: 21-Sep-16

Protein chain

Q8IXI2  (MIRO1_HUMAN) -  Mitochondrial Rho GTPase 1 from Homo sapiens

Seq: 618 a.a.

Struc: 397 a.a.

Enzyme reactions

• Enzyme class: E.C.3.6.5.- - ?????

DOI no: 10.1038/srep33019

Sci Rep 6:33019 (2016)

PubMed id: 27605430

Structural insights into Parkin substrate lysine targeting from minimal Miro substrates.

J.L.Klosowiak, S.Park, K.P.Smith, M.E.French, P.J.Focia, D.M.Freymann, S.E.Rice.

ABSTRACT

Hereditary Parkinson's disease is commonly caused by mutations in the protein kinase PINK1 or the E3 ubiquitin ligase Parkin, which function together to eliminate damaged mitochondria. PINK1 phosphorylates both Parkin and ubiquitin to stimulate ubiquitination of dozens of proteins on the surface of the outer mitochondrial membrane. However, the mechanisms by which Parkin recognizes specific proteins for modification remain largely unexplored. Here, we show that the C-terminal GTPase (cGTPase) of the Parkin primary substrate human Miro is necessary and sufficient for efficient ubiquitination. We present several new X-ray crystal structures of both human Miro1 and Miro2 that reveal substrate recognition and ubiquitin transfer to be specific to particular protein domains and lysine residues. We also provide evidence that Parkin substrate recognition is functionally separate from substrate modification. Finally, we show that prioritization for modification of a specific lysine sidechain of the cGTPase (K572) within human Miro1 is dependent on both its location and chemical microenvironment. Activation of Parkin by phosphorylation or by binding of pUb is required for prioritization of K572 for modification, suggesting that Parkin activation and acquisition of substrate specificity are coupled.