Pujals Sílvia, Bastús Neus G, Pereiro Eva, López-Iglesias Carmen, Puntes Víctor F, Kogan Marcelo J, Giralt Ernest
Institut de Recerca Biomèdica, Parc Científic de Barcelona, Spain.
Chembiochem. 2009 Apr 17;10(6):1025-31. doi: 10.1002/cbic.200800843.
Cell-penetrating peptides (CPPs) are a potential tool for intracellular delivery of different kinds of cargoes. Because of their growing use in nanobiomedicine, both for diagnostics and for treatment, metal nanoparticles are an interesting cargo for CPPs. Here, gold nanoparticles (AuNps) and the amphipathic proline-rich peptide SAP have been used. Conjugation of the peptide onto the AuNps was achieved by addition of a cysteine to the SAP sequence for thiol chemisorption on gold, and the attachment was confirmed by visible spectroscopy, dynamic light scattering (DLS), zeta-potential (ZP), stability towards ionic strength (as high as 1 M NaCl), X-ray photoelectron spectroscopy (XPS) and high-resolution transmission electron microscopy (HR-TEM) coupled to electron energy loss spectroscopy (EELS). AuNp-C-SAP internalization in HeLa cells was observed by three different microscopy techniques-TEM, confocal laser scanning microscopy (CLSM) and transmission X-ray microscopy (TXM)-and all of them have confirmed the effective intracellular delivery of AuNps by SAP.
细胞穿透肽(CPPs)是一种用于向细胞内递送各种货物的潜在工具。由于其在纳米生物医学中用于诊断和治疗的应用日益广泛,金属纳米颗粒是CPPs的一种有趣的货物。在此,使用了金纳米颗粒(AuNps)和富含两亲性脯氨酸的肽SAP。通过在SAP序列中添加半胱氨酸以在金上进行硫醇化学吸附,实现了肽与AuNps的缀合,并通过可见光谱、动态光散射(DLS)、zeta电位(ZP)、对离子强度(高达1 M NaCl)的稳定性、X射线光电子能谱(XPS)以及与电子能量损失谱(EELS)耦合的高分辨率透射电子显微镜(HR-TEM)对附着进行了确认。通过三种不同的显微镜技术——透射电子显微镜(TEM)、共聚焦激光扫描显微镜(CLSM)和透射X射线显微镜(TXM)观察了AuNp-C-SAP在HeLa细胞中的内化,并且所有这些技术都证实了SAP能够有效地将AuNps递送至细胞内。