Time to export NTF protein through NPC (Nuclear pore complex)

Value 2.2 sec Range: ±0.2 sec
Organism Human Homo sapiens
Reference Siebrasse JP, Peters R. Rapid translocation of NTF2 through the nuclear pore of isolated nuclei and nuclear envelopes. EMBO Rep. 2002 Sep3(9):887-92. p. 890 right column top paragraphPubMed ID12189172
Method FRAP: To measure export kinetics in nuclear envelopes more precisely, researchers employed photobleaching (Keminer and Peters, 1999). After the addition of transport media containing either 15 µM GFP or 7.4 µM Alexa488 NTF2 dimers to a filter-attached nuclear envelope, the transport substrate was allowed to equilibrate between the OSTR chamber and filter pores. Then, employing the time-lapse/bleaching program of the confocal microscope system together with a high-power objective (100-fold, 1.3 numerical aperture), a prebleach scan was acquired, a filter pore selected for 1 s of photobleaching, and the recovery of the fluorescence of the photobleached filter pore followed by a series of scans.
Comments Transport times of 2.2 and 1.6 sec correspond to transport rates of 670-910 molecules/NPC/Sec (?C=7.4µM) and 91-123 molecules/NPC/Sec (?C=1µM)
Entered by Uri M
ID 105653