relationships between membrane binding, affinity and cell internalization efficacy of a cell-penetrating peptide penetratin as a case study膜之间的关系绑定,亲和力和细胞内化的效果cell-penetrating肽penetratin作为案例研究.pdfVIP

Relationships between Membrane Binding, Affinity and Cell Internalization Efficacy of a Cell-Penetrating Peptide: Penetratin as a Case Study 1,2,3 ¤ 1,2,3 1,2,3 1,2,3 1,2,3 Isabel D. Alves * , Cherine Bechara , Astrid Walrant , Yefim Zaltsman , Chen-Yu Jiao , Sandrine Sagan1,2,3* 1 UPMC Univ Paris 06, UMR 7203, LBM, Paris, France, 2 CNRS, UMR 7203, LBM, Paris, France, 3 ENS, UMR 7203, LBM, Paris, France Abstract Background: Penetratin is a positively charged cell-penetrating peptide (CPP) that has the ability to bind negatively charged membrane components, such as glycosaminoglycans and anionic lipids. Whether this primary interaction of penetratin with these cell surface components implies that the peptide will be further internalized is not clear. Methodology: Using mass spectrometry, the amount of internalized and membrane bound penetratin remaining after washings, were quantified in three different cell lines: wild type (WT), glycosaminoglycans- (GAGneg) and sialic acid-deficient (SAneg) cells. Additionally, the affinity and kinetics of the interaction of penetratin to membrane models composed of pure lipids and membrane fragments from the referred cell lines was investigated, as well as the thermodynamics of such interactions using plasmon resonance and calorimetry. Principal Findings: Penetratin internalized with the same efficacy in the three cell lines at 1 mM, but was better internalized at 10 mM in SAneg.WT.GAGneg. The heat released by the interaction of penetratin with these cells followed the ranking order of internalization efficiency. Penetratin had an affinity of 10 nM for WT cells and mM for SAneg and GAGneg cells and model membrane