1ASK

NUCLEAR TRANSPORT FACTOR 2 (NTF2) H66A MUTANT

1ASK の概要

• エントリーDOI: 10.2210/pdb1ask/pdb
• 分子名称: NUCLEAR TRANSPORT FACTOR 2 (2 entities in total)
• 機能のキーワード: transport, nuclear transport protein
• 由来する生物種: Rattus norvegicus (Norway rat)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 28848.71

構造登録者

Mccoy, A.J.,Stewart, M.J. (登録日: 1997-08-11, 公開日: 1998-01-28, 最終更新日: 2024-05-22)

主引用文献

Clarkson, W.D.,Corbett, A.H.,Paschal, B.M.,Kent, H.M.,McCoy, A.J.,Gerace, L.,Silver, P.A.,Stewart, M.
Nuclear protein import is decreased by engineered mutants of nuclear transport factor 2 (NTF2) that do not bind GDP-Ran.
J.Mol.Biol., 272:716-730, 1997

PubMed Abstract: Nuclear transport factor 2 (NTF2) is associated with the translocation stage of nuclear protein import and binds both to nuclear pore proteins (nucleoporins) containing phenylalanine-rich repeats and to the Ras family GTPase Ran. In this study we probed the role of the NTF2-Ran interaction in nuclear protein import using site-directed mutants of NTF2 that interfere with its interaction with GDP-Ran. The design of these mutants was based on the X-ray crystal structure of NTF2 and was concentrated on conserved residues in and around the molecule's hydrophobic cavity. The mutant NTF2 cDNAs were expressed in Escherichia coli. Purified mutant proteins retained the interaction with FxFG-repeat nucleoporins, but several mutants in the negatively charged residues that surround the NTF2 cavity or in residues in the cavity itself were unable to bind GDP-Ran in vitro. The crystal structure of the E42K mutant protein showed significant structural changes only in this side-chain, indicating that it participated directly in the interaction with GDP-Ran. In permeabilised cell nuclear protein import assays, only wild-type NTF2 and mutants that bound GDP-Ran were functional. Furthermore, when the NTF2 E42K and D92N/D94N NTF2 mutants that failed to bind GDP-Ran in vitro were substituted for the chromosomal yeast NTF2, the yeast cells became non-viable, whereas yeast substituted with wild-type human NTF2 remained viable. We conclude that interaction between NTF2 and GDP-Ran is important for efficient nuclear protein import.

PubMed: 9368653
DOI: 10.1006/jmbi.1997.1255

実験手法

X-RAY DIFFRACTION (2.3 Å)