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Now Available from Sigma-RBI
Prod. No. N 4160
NTF2 mediates the nuclear import of Ran-GDP, driving the nuclear accumulation
of Ran. This process is essential for maintaining cellular nuclear transport and for cell viability.
NTF2, which possesses non-overlapping binding sites for both Ran-GDP and nucleoporin FxFG repeats, binds
Ran-GDP in the cytoplasm and transports it via the nuclear pore complex (NPC) by binding to the FxFG
repeats. The NTF2-[Ran-GDP] complex transport via the NPC is facilitated by the very low affinity of NTF2
to the FxFG repeats, which allows rapid attachment and detachment of NTF2 to the nucleoporins, while it
passes through the NPC. At the nucleus side of the NPC, release of Ran-GDP from NTF2 may be promoted by
dissociation of NTF2 into monomers.
Nuclear Transport Factor 2 (NTF2, N4160) is a human, recombinant protein expressed in E. coli. It
is a homodimeric protein containing 127 amino acids and the molecular weight of the monomer is
approximately 15 kDa.
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NTF2 (Prod. No.
N4160) was added to Ran (Prod. No.
R3152) immobilized
on His-Select HC Nickel Affinity Gel (Prod. No.
P6611) (Lanes 3 & 4) or to His-Select HC Nickel
Affinity Gel alone (Lane 1 control). Assay buffer only was added to His-Select HC Nickel Affinity
Gel immobilized Ran (Lane 2 control). Following 1hr mixing at RT, the resin was washed, heated in
sample buffer and the proteins were separated on 20% SDS-PAGE.
The results show that:
Lane 1: NTF2 does not bind nonspecifically to His-Select HC Nickel Affinity Gel.
Lane 2: Ran does not show any contamination at 14-15kDa.
Lanes 3 & 4: NTF2 binds specifically to Ran. |
References
- Paschal, B.M., and Gerace, L., Identification of NTF2, a cytosolic factor for nuclear import
that interacts with nuclear pore complex protein p62. J. Cell Biol., 129, 925-937 (1995).
- Stewart, M., Insights into the molecular mechanism of nuclear trafficking
using nuclear transport factor 2 (NTF2). Cell Struct. Func., 25, 217-225 (2000).
- Quimby, B.B. et al., Functional analysis of the hydrophobic patch on nuclear transport
factor 2 involved in interactions with the nuclear pore in vivo. J. Biol. Chem., 276,
38820-38829 (2001).
- Steggerda, S.M., and Paschal, B.M., Regulation of nuclear import and export by the GTPase Ran.
Inv. Rev. Cytol., 217, 41-91 (2002).
- Chaillan-Huntington, C. et al., NTF2 monomer-dimer equilibrium. J. Mol. Biol., 314,
465-477 (2002).
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