Competitive Microtubule Binding of PEX14 Coordinates Peroxisomal Protein Import and Motility

Maren Reuter; Hamed Kooshapur; Jeff-Gordian Suda; Stefan Gaussmann; Alexander Neuhaus; Lena Brühl; Pratima Bharti; Martin Jung; Wolfgang Schliebs; Michael Sattler; Ralf Erdmann

doi:10.1016/j.jmb.2020.166765

Competitive Microtubule Binding of PEX14 Coordinates Peroxisomal Protein Import and Motility

J Mol Biol. 2021 Mar 5;433(5):166765. doi: 10.1016/j.jmb.2020.166765. Epub 2021 Jan 21.

Authors

Affiliations

¹ Institute for Biochemistry and Pathobiochemistry, Department of Systems Biology, Faculty of Medicine, Ruhr University of Bochum, 44780 Bochum, Germany.
² Munich Center for Integrated Protein Science at Chair of Biomolecular NMR, Department Chemie, Technische Universität München, Lichtenbergstr. 4, 85747 Garching, Germany; Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
³ Saarland University, 66421 Homburg, Germany.
⁴ Institute for Biochemistry and Pathobiochemistry, Department of Systems Biology, Faculty of Medicine, Ruhr University of Bochum, 44780 Bochum, Germany. Electronic address: wolfgang.schliebs@rub.de.
⁵ Munich Center for Integrated Protein Science at Chair of Biomolecular NMR, Department Chemie, Technische Universität München, Lichtenbergstr. 4, 85747 Garching, Germany; Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany. Electronic address: sattler@helmholtz-muenchen.de.
⁶ Institute for Biochemistry and Pathobiochemistry, Department of Systems Biology, Faculty of Medicine, Ruhr University of Bochum, 44780 Bochum, Germany. Electronic address: ralf.erdmann@rub.de.

PMID: 33484719
DOI: 10.1016/j.jmb.2020.166765

Abstract

Human PEX14 plays a dual role as docking protein in peroxisomal protein import and as peroxisomal anchor for microtubules (MT), which relates to peroxisome motility. For docking, the conserved N-terminal domain of PEX14 (PEX14-NTD) binds amphipathic alpha-helical ligands, typically comprising one or two aromatic residues, of which human PEX5 possesses eight. Here, we show that the PEX14-NTD also binds to microtubular filaments in vitro with a dissociation constant in nanomolar range. PEX14 interacts with two motifs in the C-terminal region of human ß-tubulin. At least one of the binding motifs is in spatial proximity to the binding site of microtubules (MT) for kinesin. Both PEX14 and kinesin can bind to MT simultaneously. Notably, binding of PEX14 to tubulin can be prevented by its association with PEX5. The data suggest that PEX5 competes peroxisome anchoring to MT by occupying the ß-tubulin-binding site of PEX14. The competitive correlation of matrix protein import and motility may facilitate the homogeneous dispersion of peroxisomes in mammalian cells.

Keywords: PEX14 binding motifs; PEX5 interaction; cytoskeleton; kinesin motor domain; peroxisome tethering.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Binding Sites
Binding, Competitive
Biological Transport
Cell Line
Escherichia coli / genetics
Escherichia coli / metabolism
Fibroblasts / cytology
Fibroblasts / metabolism
Gene Expression
Humans
Kinesins / genetics
Kinesins / metabolism
Membrane Proteins / chemistry*
Membrane Proteins / genetics
Membrane Proteins / metabolism
Microtubules / metabolism*
Models, Molecular
Peroxisome-Targeting Signal 1 Receptor / chemistry*
Peroxisome-Targeting Signal 1 Receptor / genetics
Peroxisome-Targeting Signal 1 Receptor / metabolism
Peroxisomes / metabolism*
Prohibitins
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Repressor Proteins / chemistry*
Repressor Proteins / genetics
Repressor Proteins / metabolism
Sequence Alignment
Sequence Homology, Amino Acid
Signal Transduction
Tubulin / chemistry*
Tubulin / genetics
Tubulin / metabolism

Substances

Membrane Proteins
PEX14 protein, human
PEX5 protein, human
Peroxisome-Targeting Signal 1 Receptor
Prohibitins
Recombinant Proteins
Repressor Proteins
Tubulin
Kinesins