[The characteristics of the transport into the nucleus of conjugates with normal and mutant nucleophilic sequences].
A study was made of the transport of conjugates of a fluorescently labeled protein and a synthetic peptide, corresponding to the nuclear localization sequence of the SV 40 large T-antigen, into the nuclei of cultured human (HeLa and A431) and murine (HER14) cells. A possibility for such conjugates to be transported into the nuclei of digitonin-permeabilized cells, without addition of exogenous cytosol, was demonstrated. A quantitative comparison of the transport levels of constructions with normal or altered (K128T) peptide sequence was performed, and a low selectivity of nuclear transport with this alteration in the digitonin-permeabilized cell system was revealed, whereas constructions with the mutant sequence, when injected into live cells, remained in the cytoplasm. ATP-dependent transport of constructions with the mutant sequence into permeabilized cell nuclei was demonstrated, with a considerable part of this transport being NEM-insensitive. A suggestion is put forward that there are several variants of the nucleophilic sequence containing protein transport in the digitonin-permeabilized cell system.
Barbul
A I
AI
Goriunov
D E
DE
Maĭboroda
O A
OA
Vinogradova
N A
NA
Skovorodkin
I N
IN
Nikol'skiĭ
N N
NN
D016475:3T3 Cells; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism*; D001692:Biological Transport; D002463:Cell Membrane Permeability / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism*; D004072:Digitonin / Q000494:pharmacology; D006801:Humans; D051379:Mice; D008845:Microinjections; D009154:Mutation / Q000502:physiology*; D021382:Protein Sorting Signals / Q000493:pharmacokinetics*; D012709:Serum Albumin / Q000493:pharmacokinetics*; D013539:Simian virus 40 / Q000276:immunology*; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D000952:Antigens, Polyomavirus Transforming; D021382:Protein Sorting Signals; D012709:Serum Albumin; D004072:Digitonin
Kharakteristika transporta v iadro kon''iugatov s normal'noĭ i mutantnoĭ nukleofil'nymi posledovatel'nostiami.