Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans
Artikel i vetenskaplig tidskrift, 2012

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.

protein

human immunodeficiency virus

membrane interactions

sialic

Penetratin

acids

macropinocytosis

Proteoglycan

delivery

Cellular uptake

enters cells

hiv-1 tat

glycosaminoglycans

endocytosis

Arginine

Membrane affinity

Cell-penetrating peptide

Författare

Helene Åmand

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Hanna Rydberg

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Louise Fornander

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Per Lincoln

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Bengt Nordén

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Elin Esbjörner Winters

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Biochimica et Biophysica Acta - Biomembranes

0005-2736 (ISSN)

Vol. 1818 11 2669-2678

Ämneskategorier

Fysikalisk kemi

Styrkeområden

Energi

Livsvetenskaper och teknik

Materialvetenskap

DOI

10.1016/j.bbamem.2012.06.006

Mer information

Skapat

2017-10-07