Cell-penetrating Peptides A REEVALUATION OF THE MECHANISM OF CELLULAR UPTAKE

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作者：

JP Richard，K Melikov，E Vives，C Ramos，B Lebleu

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摘要：

Cellular uptake of a family of cationic cell-penetrating (examples include Tat and penetratin) have been ascribed in the literature to a mechanism that does not involve . In this work we reevaluate the mechanisms of cellular uptake of Tat 48-60 and ()(9). We demonstrate here that cell fixation, even in mild conditions, leads to the artifactual uptake of these . Moreover, we show that flow cytometry analysis cannot be used validly to evaluate cellular uptake unless a step of of the -adsorbed is included in the protocol. Fluorescence microscopy on live unfixed cells shows characteristic endosomal distribution of . Flow cytometry analysis indicates that the kinetics of uptake are similar to the kinetics of . uptake is inhibited by incubation at low temperature and cellular ATP pool depletion. Similar data were obtained for Tat-conjugated . These data are consistent with the involvement of in the cellular internalization of cell-penetrating and their conjugates to .

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关键词：

Cell Line Animals Humans Trypsin Peptides Gene Products, tat Adenosine Triphosphate Fixatives Fluorescent Dyes Microscopy, Fluorescence