Direct peptide interaction with surface glycosaminoglycans contributes to the cell penetration of maurocalcine. - Grenoble Institut des Neurosciences Accéder directement au contenu
Article Dans Une Revue Journal of Biological Chemistry Année : 2008

Direct peptide interaction with surface glycosaminoglycans contributes to the cell penetration of maurocalcine.

Résumé

Maurocalcine (MCa), initially identified from a Tunisian scorpion venom, defines a new member of the family of cell penetrating peptides by its ability to efficiently cross the plasma membrane. The initiating mechanistic step required for the cell translocation of a cell penetrating peptide implicates its binding onto cell surface components such as membrane lipids and/or heparan sulfate proteoglycans. Here we characterized the interaction of wild-type MCa and MCa K20A, a mutant analogue with reduced cell-penetration efficiency, with heparin (HP) and heparan sulfates (HS) through surface plasma resonance. HP and HS bind both to MCa, indicating that heparan sulfate proteoglycans may represent an important entry route of the peptide. This is confirmed by the fact that (i) both compounds bind with reduced affinity to MCa K20A and (ii) the cell penetration of wild-type or mutant MCa coupled to fluorescent streptavidin is reduced by about 50% in mutant Chinese hamster ovary cell lines lacking either all glycosaminoglycans (GAGs) or just HS. Incubating MCa with soluble HS, HP, or chondroitin sulfates also inhibits the cell penetration of MCa-streptavidin complexes. Analyses of the cell distributions of MCa/streptavidin in several Chinese hamster ovary cell lines show that the distribution of the complex coincides with the endosomal marker Lyso-Tracker red and is not affected by the absence of GAGs. The distribution of MCa/streptavidin is not coincident with that of transferrin receptors nor affected by a dominant-negative dynamin 2 K44A mutant, an inhibitor of clathrin-mediated endocytosis. However, entry of the complex is greatly diminished by amiloride, indicating the importance of macropinocytosis in MCa/streptavidin entry. It is concluded that (i) interaction of MCa with GAGs quantitatively improves the cell penetration of MCa, and (ii) GAG-dependent and -independent MCa penetration rely similarly on the macropinocytosis pathway.
Fichier principal
Vignette du fichier
inserm-00376511_edited.pdf (2.31 Mo) Télécharger le fichier
GAGS-MCa-RAM-2008-ver2-FIG.pdf (963.94 Ko) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)
Origine : Fichiers produits par l'(les) auteur(s)
Loading...

Dates et versions

inserm-00376511 , version 1 (29-04-2009)

Identifiants

Citer

Narendra Ram, Sonia Aroui, Emilie Jaumain, Hicham Bichraoui, Kamel Mabrouk, et al.. Direct peptide interaction with surface glycosaminoglycans contributes to the cell penetration of maurocalcine.. Journal of Biological Chemistry, 2008, 283 (35), pp.24274-84. ⟨10.1074/jbc.M709971200⟩. ⟨inserm-00376511⟩
279 Consultations
395 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More