核定位信号与质粒DNA的偶联以及偶联物与输入蛋白α的特异性相互作用。

Coupling of nuclear localization signals to plasmid DNA and specific interaction of the conjugates with importin alpha.

作者信息

Ciolina C, Byk G, Blanche F, Thuillier V, Scherman D, Wils P

机构信息

UMR 133 CNRS/Rhône-Poulenc Rorer and Rhône-Poulenc Rorer Gencell, Centre de Recherche de Vitry Alfortville, B.P.14, 13 quai Jules Guesde, 94403 Vitry-sur-Seine Cedex, France.

出版信息

Bioconjug Chem. 1999 Jan-Feb;10(1):49-55. doi: 10.1021/bc980061a.

DOI:10.1021/bc980061a

PMID:9893963

Abstract

The nuclear localization signal (NLS) of the SV40 large T antigen efficiently induces nuclear targeting of proteins. We have developed a chemical strategy for covalent coupling of NLS peptides to plasmid DNA. A p-azido-tetrafluoro-benzyl-NLS peptide conjugate was synthesized. This conjugate was used to covalently associate NLS peptides to plasmid DNA by photoactivation. Reporter gene was expressed after transfection of the plasmid-NLS conjugates in NIH 3T3 cells. The conjugates interacted specifically with the NLS-receptor importin alpha, but plasmid-NLS conjugates were not detected in the nucleus, by fluorescence microscopy, after cytoplasmic microinjection.

摘要

猴空泡病毒40大T抗原的核定位信号（NLS）能有效诱导蛋白质的核靶向。我们已开发出一种将NLS肽与质粒DNA共价偶联的化学策略。合成了一种对叠氮基四氟苄基-NLS肽共轭物。该共轭物通过光活化用于将NLS肽与质粒DNA共价结合。在将质粒-NLS共轭物转染到NIH 3T3细胞后，报告基因得以表达。这些共轭物与NLS受体输入蛋白α特异性相互作用，但通过荧光显微镜观察，在细胞质显微注射后，未在细胞核中检测到质粒-NLS共轭物。