Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Biophys J
2003 Oct 01;854:2311-22. doi: 10.1016/S0006-3495(03)74655-4.
Show Gene links
Show Anatomy links
Optical microwell assay of membrane transport kinetics.
Kiskin NI
,
Siebrasse JP
,
Peters R
.
???displayArticle.abstract???
In optical single transporter recording, membranes are firmly attached to flat solid substrates containing small wells or test compartments (TC). Transport of fluorescent molecules through TC-spanning membrane patches is induced by solution change and recorded by confocal microscopy. Previously, track-etched membrane filters were used to create solid substrates containing populations of randomly distributed TCs. In this study the possibilities offered by orderly TC arrays as created by laser microdrilling were explored. A theoretical framework was developed taking the convolution of membrane transport, solution change, and diffusion into account. The optical properties of orderly TC arrays were studied and the kinetics of solution change measured. Export and import through the nuclear pore complex (NPC) was analyzed in isolated envelopes of Xenopus oocyte nuclei. In accordance with previous reports nuclear transport receptor NTF2, which binds directly to NPC proteins, was found to be translocated much faster than "inert" molecules of similar size. Unexpectedly, NXT1, a homolog of NTF2 reportedly unable to bind to NPC proteins directly, was translocated as fast as NTF2. Thus, microstructured TC arrays were shown to provide optical single transporter recording with a new basis.
Adamson,
Single capillary permeability to proteins having similar size but different charge.
1988, Pubmed
Adamson,
Single capillary permeability to proteins having similar size but different charge.
1988,
Pubmed
Allen,
The nuclear pore complex: mediator of translocation between nucleus and cytoplasm.
2000,
Pubmed
Bayliss,
Structural basis for the interaction between NTF2 and nucleoporin FxFG repeats.
2002,
Pubmed
Black,
Identification of an NTF2-related factor that binds Ran-GTP and regulates nuclear protein export.
1999,
Pubmed
Black,
NXT1 is necessary for the terminal step of Crm1-mediated nuclear export.
2001,
Pubmed
Braun,
Overexpression of TAP/p15 heterodimers bypasses nuclear retention and stimulates nuclear mRNA export.
2001,
Pubmed
,
Xenbase
Braun,
Nuclear export of mRNA by TAP/NXF1 requires two nucleoporin-binding sites but not p15.
2002,
Pubmed
,
Xenbase
Bullock,
The 1.6 angstroms resolution crystal structure of nuclear transport factor 2 (NTF2).
1996,
Pubmed
Chaillan-Huntington,
NTF2 monomer-dimer equilibrium.
2001,
Pubmed
,
Xenbase
Chaillan-Huntington,
Dissecting the interactions between NTF2, RanGDP, and the nucleoporin XFXFG repeats.
2000,
Pubmed
Clarkson,
Nuclear protein import is decreased by engineered mutants of nuclear transport factor 2 (NTF2) that do not bind GDP-Ran.
1997,
Pubmed
Dworetzky,
The effects of variations in the number and sequence of targeting signals on nuclear uptake.
1988,
Pubmed
,
Xenbase
Fahrenkrog,
Nuclear pore complex architecture and functional dynamics.
2001,
Pubmed
Fribourg,
Structural basis for the recognition of a nucleoporin FG repeat by the NTF2-like domain of the TAP/p15 mRNA nuclear export factor.
2001,
Pubmed
Grote,
Mapping of nucleoporins to the center of the nuclear pore complex by post-embedding immunogold electron microscopy.
1995,
Pubmed
Guzik,
NXT1 (p15) is a crucial cellular cofactor in TAP-dependent export of intron-containing RNA in mammalian cells.
2001,
Pubmed
Katahira,
The Mex67p-mediated nuclear mRNA export pathway is conserved from yeast to human.
1999,
Pubmed
Keminer,
Permeability of single nuclear pores.
1999,
Pubmed
,
Xenbase
Keminer,
Optical recording of signal-mediated protein transport through single nuclear pore complexes.
1999,
Pubmed
,
Xenbase
Kiskin,
Two-photon excitation and photolysis by pulsed laser illumination modelled by spatially non-uniform reactions with simultaneous diffusion.
2002,
Pubmed
Lévesque,
RNA export mediated by tap involves NXT1-dependent interactions with the nuclear pore complex.
2001,
Pubmed
Macara,
Transport into and out of the nucleus.
2001,
Pubmed
Moore,
Purification of a Ran-interacting protein that is required for protein import into the nucleus.
1994,
Pubmed
,
Xenbase
Nehrbass,
Role of the nuclear transport factor p10 in nuclear import.
1996,
Pubmed
Ossareh-Nazari,
RanGTP-binding protein NXT1 facilitates nuclear export of different classes of RNA in vitro.
2000,
Pubmed
,
Xenbase
Paine,
Nuclear envelope permeability.
1975,
Pubmed
Panté,
Nuclear pore complex is able to transport macromolecules with diameters of about 39 nm.
2002,
Pubmed
,
Xenbase
Paschal,
Identification of NTF2, a cytosolic factor for nuclear import that interacts with nuclear pore complex protein p62.
1995,
Pubmed
Peters,
Optical single transporter recording: transport kinetics in microarrays of membrane patches.
2003,
Pubmed
Peters,
Nucleo-cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis.
1984,
Pubmed
Peters,
Fluorescence microphotolysis to measure nucleocytoplasmic transport and intracellular mobility.
1986,
Pubmed
Ribbeck,
Kinetic analysis of translocation through nuclear pore complexes.
2001,
Pubmed
Ribbeck,
NTF2 mediates nuclear import of Ran.
1998,
Pubmed
Rout,
The nuclear pore complex as a transport machine.
2001,
Pubmed
Rout,
The yeast nuclear pore complex: composition, architecture, and transport mechanism.
2000,
Pubmed
Ryan,
The nuclear pore complex: a protein machine bridging the nucleus and cytoplasm.
2000,
Pubmed
Siebrasse,
Reconstitution of nuclear protein export in isolated nuclear envelopes.
2002,
Pubmed
,
Xenbase
Siebrasse,
Rapid translocation of NTF2 through the nuclear pore of isolated nuclei and nuclear envelopes.
2002,
Pubmed
,
Xenbase
Smith,
Nuclear import of Ran is mediated by the transport factor NTF2.
,
Pubmed
Stewart,
Molecular mechanism of translocation through nuclear pore complexes during nuclear protein import.
2001,
Pubmed
Stewart,
Structural basis for molecular recognition between nuclear transport factor 2 (NTF2) and the GDP-bound form of the Ras-family GTPase Ran.
1998,
Pubmed
Suyama,
Prediction of structural domains of TAP reveals details of its interaction with p15 and nucleoporins.
2000,
Pubmed
Swaminathan,
Photobleaching recovery and anisotropy decay of green fluorescent protein GFP-S65T in solution and cells: cytoplasmic viscosity probed by green fluorescent protein translational and rotational diffusion.
1997,
Pubmed
Tschödrich-Rotter,
An optical method for recording the activity of single transporters in membrane patches.
1998,
Pubmed
Vasu,
Nuclear pores and nuclear assembly.
2001,
Pubmed
