Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans

Handle URI:
http://hdl.handle.net/10754/597743
Title:
Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans
Authors:
Åmand, Helene L.; Rydberg, Hanna A.; Fornander, Louise H.; Lincoln, Per; Nordén, Bengt; Esbjörner, Elin K.
Abstract:
Cell 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.
Citation:
Åmand 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.
Publisher:
Elsevier BV
Journal:
Biochimica et Biophysica Acta (BBA) - Biomembranes
Issue Date:
Nov-2012
DOI:
10.1016/j.bbamem.2012.06.006
PubMed ID:
22705501
Type:
Article
ISSN:
0005-2736
Sponsors:
Prof. 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.
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Full metadata record

DC FieldValue Language
dc.contributor.authorÅmand, Helene L.en
dc.contributor.authorRydberg, Hanna A.en
dc.contributor.authorFornander, Louise H.en
dc.contributor.authorLincoln, Peren
dc.contributor.authorNordén, Bengten
dc.contributor.authorEsbjörner, Elin K.en
dc.date.accessioned2016-02-25T12:55:55Zen
dc.date.available2016-02-25T12:55:55Zen
dc.date.issued2012-11en
dc.identifier.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.en
dc.identifier.issn0005-2736en
dc.identifier.pmid22705501en
dc.identifier.doi10.1016/j.bbamem.2012.06.006en
dc.identifier.urihttp://hdl.handle.net/10754/597743en
dc.description.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.en
dc.description.sponsorshipProf. 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.en
dc.publisherElsevier BVen
dc.subjectArginineen
dc.subjectCell-penetrating peptideen
dc.subjectCellular uptakeen
dc.subjectMembrane affinityen
dc.subjectPenetratinen
dc.subjectProteoglycanen
dc.titleCell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycansen
dc.typeArticleen
dc.identifier.journalBiochimica et Biophysica Acta (BBA) - Biomembranesen
dc.contributor.institutionChalmers University of Technology, Göteborg, Swedenen

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