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使用凝集素和聚乙二醇使含糖脂的脂质体与真核细胞融合。

Use of lectins and polyethylene glycol for fusion of glycolipid-containing liposomes with eukaryotic cells.

作者信息

Szoka F, Magnusson K E, Wojcieszyn J, Hou Y, Derzko Z, Jacobson K

出版信息

Proc Natl Acad Sci U S A. 1981 Mar;78(3):1685-9. doi: 10.1073/pnas.78.3.1685.

Abstract

Efficient fusion of phospholipid vesicles with monolayer cultures of eukaryotic cells was accomplished by attaching glycolipid-containing vesicles to the cell surface by using a lectin displaying binding for both the cell surface and the glycolipid, followed by treatment with polyethylene glycol. Fusion was inferred from the transfer of fluorescent lipid analog probes embedded in the vesicle membrane over the entire cell surface and of fluoresceinated proteins from the aqueous space of the vesicle to the cytoplasm of the cell. Fluorescence recovery after photobleaching showed that both the injected membrane and the cytoplasmic markers were mobile. Two different lectin--glycolipid combinations [Ricinus communis agglutinin I-lactosylcerebroside and concanavalin A-tetradecyl- (or hexadecyl-) maltobionamide] were used to promote attachment of lipid vesicles before polyethylene glycol-induced fusion with BG-9 human fibroblasts, NIL-8M2 hamster cells, or L-929 mouse cells. In the absence of lectin or polyethylene glycol, fusion was negligible. However, when both the lectin and the glycol were used, a dramatic increase in the transfer of both vesicle membrane and aqueous space markers from the liposomes to the cells occurred.

摘要

通过使用对细胞表面和糖脂均具有结合能力的凝集素,将含糖脂的囊泡附着于细胞表面,随后用聚乙二醇处理,实现了磷脂囊泡与真核细胞单层培养物的高效融合。融合现象可通过嵌入囊泡膜的荧光脂质类似物探针在整个细胞表面的转移以及荧光蛋白从囊泡水相空间转移至细胞胞质来推断。光漂白后的荧光恢复表明,注入的膜标记物和细胞质标记物均具有流动性。在聚乙二醇诱导脂质囊泡与BG - 9人成纤维细胞、NIL - 8M2仓鼠细胞或L - 929小鼠细胞融合之前,使用了两种不同的凝集素 - 糖脂组合[蓖麻凝集素I - 乳糖基神经酰胺和伴刀豆球蛋白A - 十四烷基 - (或十六烷基) - 麦芽糖二酰胺]来促进脂质囊泡的附着。在没有凝集素或聚乙二醇的情况下,融合可忽略不计。然而,当同时使用凝集素和糖脂时,脂质体膜标记物和水相空间标记物从脂质体向细胞的转移显著增加。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3293/319197/caf9a709adb1/pnas00654-0400-a.jpg

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