3FF5

Crystal structure of the conserved N-terminal domain of the peroxisomal matrix-protein-import receptor, Pex14p

Summary for 3FF5

• Entry DOI: 10.2210/pdb3ff5/pdb
• Descriptor: Peroxisomal biogenesis factor 14, decyl 2-trimethylazaniumylethyl phosphate (3 entities in total)
• Functional Keywords: protein import, peroxin, 3 helices bundle, protein transport
• Biological source: Rattus norvegicus (Rat)
• Cellular location: Peroxisome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity): Q642G4
• Total number of polymer chains: 2
• Total formula weight: 12455.32

Authors

Su, J.-R.,Takeda, K.,Tamura, S.,Fujiki, Y.,Miki, K. (deposition date: 2008-12-01, release date: 2008-12-30, Last modification date: 2023-12-27)

Primary citation

Su, J.R.,Takeda, K.,Tamura, S.,Fujiki, Y.,Miki, K.
Crystal structure of the conserved N-terminal domain of the peroxisomal matrix protein import receptor, Pex14p
Proc.Natl.Acad.Sci.USA, 106:417-421, 2009

PubMed Abstract: Pex14p is a central component of the peroxisomal protein import machinery, in which the conserved N-terminal domain mediates dynamic interactions with other peroxins including Pex5p, Pex13p, and Pex19p. Here, we report the crystal structure of the conserved N-terminal domain of Pex14p with a three-helix bundle. A hydrophobic surface is composed of the conserved residues, of which two phenylalanine residues (Phe-35 and Phe-52) protrude to the solvent. Consequently, two putative binding pockets suitable for recognizing the helical WXXXF/Y motif of Pex5p are formed on the surface by the two phenylalanine residues accompanying with positively charged residues. The structural feature agrees well with our earlier findings where F35A/L36A and F52A/L53A mutants were impaired in the interactions with other peroxins such as Pex5p and Pex13p. Pex14p variants each with Phe-to-Ala mutation at positions 35, 52, and 35/52, respectively, were defective in restoring the impaired peroxisomal protein import in pex14 Chinese hamster ovary mutant ZP161 cells. Moreover, in GST pull-down assays His(6)-Pex5pL bound only to GST-Pex14p(25-70), not to any of GST-Pex14p(25-70)F35A, GST-Pex14p(25-70)F52A, and GST-Pex14p(25-70)F35A/F52A. Endogenous Pex5p was recruited to FLAG-Pex14p on peroxisomes in vivo but barely to FLAG-Pex14pF35A, FLAG-Pex14pF52A, and FLAG-Pex14pF35A/F52A. Collectively, Phe-35 and Phe-52 are essential for the Pex14p functions, including the interaction between Pex14p and Pex5p.

PubMed: 19122147
DOI: 10.1073/pnas.0808681106

Experimental method

X-RAY DIFFRACTION (1.8 Å)