Trafficking of peroxisomal targeting signal 1 (PTS1) proteins to the Leishmania glycosome is dependent on the docking of the LdPEX5 receptor to LdPEX14 on the glycosomal membrane. A combination of deletion and random mutagenesis was used to identify residues in the LdPEX14 N-terminal region that are critical for mediating the LdPEX5–LdPEX14 interaction. These studies highlighted residues 35–75 on ldpex14 as the core domain required for binding LdPEX5. Single point mutation within this core domain generally did not affect the ldpex5-(203–391)–ldpex14-(1–120) interaction; notable exceptions were substitutions at Phe40, Val46 or Phe57 which completely abolished or increased the apparent Kd value for ldpex5-(203–391) binding 30-fold. Biochemical studies revealed that these point mutations did not alter either the secondary or quaternary structure of LdPEX14 and indicated that the latter residues were critical for stabilizing the LdPEX5–LdPEX14 interaction.
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Research Article|
January 06 2015
Identification of Leishmania donovani peroxin 14 residues required for binding the peroxin 5 receptor proteins
Hamed Hojjat;
Hamed Hojjat
*Institute of Parasitology and Centre for Host-Parasite Interactions, McGill University, Ste. Anne de Bellevue, Quebec, Canada, H9X 3V9
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Armando Jardim
Armando Jardim
1
*Institute of Parasitology and Centre for Host-Parasite Interactions, McGill University, Ste. Anne de Bellevue, Quebec, Canada, H9X 3V9
1To whom correspondence should be addressed (email armando.jardim@mcgill.ca).
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Publisher: Portland Press Ltd
Received:
September 08 2014
Revision Received:
October 20 2014
Accepted:
November 05 2014
Accepted Manuscript online:
November 05 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2015 Biochemical Society
2015
Biochem J (2015) 465 (2): 247–257.
Article history
Received:
September 08 2014
Revision Received:
October 20 2014
Accepted:
November 05 2014
Accepted Manuscript online:
November 05 2014
Citation
Hamed Hojjat, Armando Jardim; Identification of Leishmania donovani peroxin 14 residues required for binding the peroxin 5 receptor proteins. Biochem J 15 January 2015; 465 (2): 247–257. doi: https://doi.org/10.1042/BJ20141133
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