Lanford R E, Kanda P, Kennedy R C
Cell. 1986 Aug 15;46(4):575-82. doi: 10.1016/0092-8674(86)90883-4.
A system was developed for the analysis of protein transport to the nucleus. Carrier proteins cross-linked to synthetic peptides were microinjected into the cytoplasm of mammalian cells, and protein transport was evaluated by immunofluorescence staining of fixed cells. A 13-mer synthetic peptide containing seven amino acids homologous to SV40 T antigen was capable of inducing nuclear transport, but no transport was observed when proteins were coupled with a synthetic peptide homologous to a nuclear-transport-defective T antigen. The largest protein-peptide conjugate efficiently transported was ferritin (Mr 465,000). The rate of transport was influenced by the number of peptides per molecule of carrier protein and, to some degree, by the size of the carrier protein. Transport of some conjugates was almost complete in 15 min at room temperature.
开发了一种用于分析蛋白质向细胞核转运的系统。将与合成肽交联的载体蛋白显微注射到哺乳动物细胞的细胞质中，并通过对固定细胞进行免疫荧光染色来评估蛋白质转运。一种含有七个与SV40 T抗原同源氨基酸的13聚体合成肽能够诱导核转运，但当蛋白质与与核转运缺陷型T抗原同源的合成肽偶联时，未观察到转运。能够有效转运的最大蛋白质-肽缀合物是铁蛋白（分子量465,000）。转运速率受每分子载体蛋白上肽的数量影响，并且在一定程度上受载体蛋白大小的影响。在室温下，一些缀合物的转运在15分钟内几乎完成。
