Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans
AuthorsÅmand, Helene L.
Rydberg, Hanna A.
Fornander, Louise H.
Esbjörner, Elin K.
Permanent link to this recordhttp://hdl.handle.net/10754/597743
MetadataShow full item record
AbstractCell surface proteoglycans (PGs) appear to promote uptake of arginine-rich cell-penetrating peptides (CPPs), but their exact functions are unclear. To address if there is specificity in the interactions of arginines and PGs leading to improved internalization, we used flow cytometry to examine uptake in relation to cell surface binding for penetratin and two arginine/lysine substituted variants (PenArg and PenLys) in wildtype CHO-K1 and PG-deficient A745 cells. All peptides were more efficiently internalized into CHO-K1 than into A745, but their cell surface binding was independent of cell type. Thus, PGs promote internalization of cationic peptides, irrespective of the chemical nature of their positive charges. Uptake of each peptide was linearly dependent on its cell surface binding, and affinity is thus important for efficiency. However, the gradients of these linear dependencies varied significantly. Thus each peptide's ability to stimulate uptake once bound to the cell surface is reliant on formation of specific uptake-promoting interactions. Heparin affinity chromatography and clustering experiments showed that penetratin and PenArg binding to sulfated sugars is stabilized by hydrophobic interactions and result in clustering, whereas PenLys only interacts through electrostatic attraction. This may have implications for the molecular mechanisms behind arginine-specific uptake stimulation as penetratin and PenArg are more efficiently internalized than PenLys upon interaction with PGs. However, PenArg is also least affected by removal of PGs. This indicates that an increased arginine content not only improve PG-dependent uptake but also that PenArg is more adaptable as it can use several portals of entry into the cell. © 2012 Elsevier B.V.
CitationÅmand HL, Rydberg HA, Fornander LH, Lincoln P, Nordén B, et al. (2012) Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans. Biochimica et Biophysica Acta (BBA) - Biomembranes 1818: 2669–2678. Available: http://dx.doi.org/10.1016/j.bbamem.2012.06.006.
SponsorsProf. Ü. Langel, Stockholm University, is acknowledged for the gift of the CHO-K1 and CHO-pgsA745 cells. This work was funded by grants to E.K.E. from the Wenner-Gren foundations and the Stenbäck foundation and to B.N. from the European Research Council and King Abdullah University of Science and Technology.
CollectionsPublications Acknowledging KAUST Support
- Stimulated endocytosis in penetratin uptake: effect of arginine and lysine.
- Authors: Amand HL, Fant K, Nordén B, Esbjörner EK
- Issue date: 2008 Jul 11
- Binding of cell-penetrating penetratin peptides to plasma membrane vesicles correlates directly with cellular uptake.
- Authors: Amand HL, Boström CL, Lincoln P, Nordén B, Esbjörner EK
- Issue date: 2011 Jul
- Delivery of antisense peptide nucleic acids to cells by conjugation with small arginine-rich cell-penetrating peptide (R/W)9.
- Authors: Cordier C, Boutimah F, Bourdeloux M, Dupuy F, Met E, Alberti P, Loll F, Chassaing G, Burlina F, Saison-Behmoaras TE
- Issue date: 2014
- Enthalpy-driven interactions with sulfated glycosaminoglycans promote cell membrane penetration of arginine peptides.
- Authors: Takechi-Haraya Y, Nadai R, Kimura H, Nishitsuji K, Uchimura K, Sakai-Kato K, Kawakami K, Shigenaga A, Kawakami T, Otaka A, Hojo H, Sakashita N, Saito H
- Issue date: 2016 Jun
- Cellular internalization and distribution of arginine-rich peptides as a function of extracellular peptide concentration, serum, and plasma membrane associated proteoglycans.
- Authors: Kosuge M, Takeuchi T, Nakase I, Jones AT, Futaki S
- Issue date: 2008 Mar