GAP43 shows partial co-localisation but no strong physical interaction with prolyl oligopeptidase

Z. Szeltner, Markus Morawski, T. Juhász, Ilona Szamosi, K. Liliom, Veronika Csizmók, F. Tölgyesi, L. Polgár

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

It has recently been proposed that prolyl oligopeptidase (POP), the cytosolic serine peptidase with neurological implications, binds GAP43 (Growth-Associated Protein 43) and is implicated in neuronal growth cone formation, axon guidance and synaptic plasticity. We investigated the interaction between GAP43 and POP with various biophysical and biochemical methods in vitro and studied the co-localisation of the two proteins in differentiated HeLa cells. GAP43 and POP showed partial co-localisation in the cell body as well as in the potential growth cone structures. We could not detect significant binding between the recombinantly expressed POP and GAP43 using gel filtration, CD, ITC and BIACORE studies, pull-down experiments, glutaraldehyde cross-linking and limited proteolysis. However, glutaraldehyde cross-linking suggested a weak and transient interaction between the proteins. Both POP and GAP43 interacted with artificial lipids in our in vitro model system, but the presence of lipids did not evoke binding between them. In native polyacrylamide gel electrophoresis, GAP43 interacted with one of the three forms of a polyhistidine-tagged prolyl oligopeptidase. The interaction of the two proteins was also evident in ELISA and we have observed co-precipitation of the two proteins during co-incubation at higher concentrations. Our results indicate that there is no strong and direct interaction between POP and GAP43 at physiological conditions.

Original languageEnglish
Pages (from-to)2162-2176
Number of pages15
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1804
Issue number12
DOIs
Publication statusPublished - Dec 2010

Fingerprint

prolyl oligopeptidase
GAP-43 Protein
Growth Cones
Glutaral
Cones
Proteins
Proteolysis
Lipids
Native Polyacrylamide Gel Electrophoresis
Neuronal Plasticity
Coprecipitation
Electrophoresis
HeLa Cells
Serine
Gel Chromatography
Plasticity
Peptide Hydrolases
Gels
Enzyme-Linked Immunosorbent Assay
Cells

Keywords

  • GAP43
  • Growth cone
  • Model lipids
  • Neuromodulin
  • Prolyl oligopeptidase
  • Protein-protein interaction

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Analytical Chemistry
  • Molecular Biology

Cite this

GAP43 shows partial co-localisation but no strong physical interaction with prolyl oligopeptidase. / Szeltner, Z.; Morawski, Markus; Juhász, T.; Szamosi, Ilona; Liliom, K.; Csizmók, Veronika; Tölgyesi, F.; Polgár, L.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1804, No. 12, 12.2010, p. 2162-2176.

Research output: Contribution to journalArticle

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