Recombinant human peroxisomal targeting signal receptor PEX5. Structural basis for interaction of PEX5 with PEX14

Wolfgang Schliebs, Jürgen Saidowsky, Bogos Agianian, Gabriele Dodt, Friedrich W. Herberg, Wolf H. Kunau

Research output: Contribution to journalArticle

134 Citations (Scopus)

Abstract

Import of matrix proteins into peroxisomes requires two targeting signal-specific import receptors, Pex5p and Pex7p, and their binding partners at the peroxisomal membrane, Pex13p and Pex14p. Several constructs of human PEX5 have been overexpressed and purified by affinity chromatography in order to determine functionally important interactions and provide initial structural information. Sizing chromatography and electron microscopy suggest that the two isoforms of the human PTS1 receptor, PEX5L and PEX5S, form homotetramers. Surface plasmon resonance analysis indicates that PEX5 binds to the N-terminal fragment of PEX14-(1-78) with a very high affinity in the low nanomolar range. Stable complexes between recombinant PEX14-(1-78) and both the full-length and truncated versions of PEX5 were formed in vitro. Analysis of these complexes revealed that PEX5 possesses multiple binding sites for PEX14, which appear to be distributed throughout its N-terminal half. Coincidentally, this part of the molecule is also responsible for oligomerization, whereas the C-terminal half with its seven tetratricopeptide repeats has been reported to bind PTS1-proteins. A pentapeptide motif that is reiterated seven times in PEX5 is proposed as a determinant for the interaction with PEX14.

Original languageEnglish (US)
Pages (from-to)5666-5673
Number of pages8
JournalJournal of Biological Chemistry
Volume274
Issue number9
DOIs
StatePublished - Feb 26 1999
Externally publishedYes

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Affinity chromatography
Oligomerization
Peroxisomes
Surface Plasmon Resonance
Surface plasmon resonance
Chromatography
Affinity Chromatography
Electron microscopy
Electron Microscopy
Protein Isoforms
Proteins
Binding Sites
Membranes
Molecules
peroxisome-targeting signal 1 receptor
peroxisomal targeting signal 2 receptor
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

Recombinant human peroxisomal targeting signal receptor PEX5. Structural basis for interaction of PEX5 with PEX14. / Schliebs, Wolfgang; Saidowsky, Jürgen; Agianian, Bogos; Dodt, Gabriele; Herberg, Friedrich W.; Kunau, Wolf H.

In: Journal of Biological Chemistry, Vol. 274, No. 9, 26.02.1999, p. 5666-5673.

Research output: Contribution to journalArticle

Schliebs, Wolfgang ; Saidowsky, Jürgen ; Agianian, Bogos ; Dodt, Gabriele ; Herberg, Friedrich W. ; Kunau, Wolf H. / Recombinant human peroxisomal targeting signal receptor PEX5. Structural basis for interaction of PEX5 with PEX14. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 9. pp. 5666-5673.
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