1,729 research outputs found
Exo70-Mediated Recruitment of Nucleoporin Nup62 at the Leading Edge of Migrating Cells is Required for Cell Migration
Nucleoporin Nup62 localizes at the central channel of the nuclear pore complex and is essential for nucleocytoplasmic transport. Through its FG-repeat domain, Nup62 regulates nuclear pore permeability and binds nuclear transport receptors. Here, we report that Nup62 interacts directly with Exo70 and colocalizes with Exo70 at the leading edge of migrating cells. Nup62 binds the N-terminal domain of Exo70 through its coiled-coil domain but not through its FG-repeat domain. Selective inhibition of leading edge Nup62 using RNA interference significantly reduces cell migration. Furthermore, Exo70 recruits Nup62 at the plasma membrane and at filopodia. Removal of the Exo70-binding domain of Nup62 prevents leading edge localization of Nup62. Analogous to Exo70, Nup62 cycles between the plasma membrane and the perinuclear recycling compartment. Altogether, we propose that Nup62 not solely regulates access to the cell nucleus, but additionally functions in conjunction with Exo70, a key regulator of exocytosis and actin dynamics, at the leading edge of migrating cells
Designer molecular probes for phosphonium ionic liquids
Investigations into the extent of structuring present in phosphonium based ionic liquids (ILs) have been carried out using photochromic molecular probes. Three spiropyran derivatives containing hydroxyl (BSP-1), carboxylic acid (BSP-2) and aliphatic chain (C14H29) (BSP-3) functional groups have been analysed in a range of phosphonium based ionic liquids and their subsequent physico-chemical interactions were reported. It is believed that the functional groups locate the probe molecules into specific regions based upon the interaction of the functional groups with particular and defined regions of the ionic liquid. This structuring results in thermodynamic, kinetic and solvatochromic parameters that are not predictable from classical solvent models. BSP-1 and BSP-2 exhibit generally negative entropies of activation ranging from -50 J K-1 mol-1 to -90 J K-1 mol-1 implying relatively low solventâsolute interactions and possible anion interactions with IL polar functional groups. Higher than expected activation energies of 60 kJ mol-1 to 100 kJ mol-1 obtained for polar probes maybe be due to IL functional groups competing with the charged sites of the merocyanine (MC) isomer thus reducing MC stabilisation effects. Differences in thermal relaxation rate constants (2.5 Ă 10-3 s-1 in BSP-1 and 3 Ă 10-4 s-1 in BSP-2 in [P6,6,6,14][dbsa]) imply that while the polar probe systems are primarily located in polar/charged regions, each probe experiences slightly differing polar domains. BSP-3 entropies of activation are positive and between 30 J K-1 mol-1 to 66 J K-1 mol-1. The association of the non-polar functional group is believed to locate the spiropyran moiety in the interfacial polar and non-polar regions. The thermal relaxation of the MC form causes solvent reorientation to accommodate the molecule as it reverts to its closed form. Slow thermal relaxation rate constants were obserevd in contrast to high activation energies (5 Ă 10-4 s-1 and 111.91 kJ mol-1 respectively, for BSP-3 in [P6,6,6,14][dbsa]). This may be due to steric effects arising from proposed nano-cavity formation by the alkyl chains in phosphonium based ILs
Simple biophysics underpins collective conformations of the intrinsically disordered proteins of the Nuclear Pore Complex
Nuclear Pore Complexes (NPCs) are key cellular transporter that control nucleocytoplasmic transport in eukaryotic cells, but its transport mechanism is still not understood. The centerpiece of NPC transport is the assembly of intrinsically disordered polypeptides, known as FG nucleoporins, lining its passageway. Their conformations and collective dynamics during transport are difficult to assess in vivo. In vitro investigations provide partially conflicting results, lending support to different models of transport, which invoke various conformational transitions of the FG nucleoporins induced by the cargo-carrying transport proteins. We show that the spatial organization of FG nucleoporin assemblies with the transport proteins can be understood within a first principles biophysical model with a minimal number of key physical variables, such as the average protein interaction strengths and spatial densities. These results address some of the outstanding controversies and suggest how molecularly divergent NPCs in different species can perform essentially the same function
The Electrostatic Screening Length in Concentrated Electrolytes Increases with Concentration
According to classical electrolyte theories interactions in dilute (low ion
density) electrolytes decay exponentially with distance, with the Debye
screening length the characteristic length-scale. This decay length decreases
monotonically with increasing ion concentration, due to effective screening of
charges over short distances. Thus within the Debye model no long-range forces
are expected in concentrated electrolytes. Here we reveal, using experimental
detection of the interaction between two planar charged surfaces across a wide
range of electrolytes, that beyond the dilute (Debye-Huuckel) regime the
screening length increases with increasing concentration. The screening lengths
for all electrolytes studied - including aqueous NaCl solutions, ionic liquids
diluted with propylene carbonate, and pure ionic liquids - collapse onto a
single curve when scaled by the dielectric constant. This non-monotonic
variation of the screening length with concentration, and its generality across
ionic liquids and aqueous salt solutions, demonstrates an important
characteristic of concentrated electrolytes of substantial relevance from
biology to energy storage.Comment: This document is the unedited authors' version of a Submitted Work
that was subsequently accepted for publication in the Journal of Physical
Chemistry Letters, copyright American Chemical Society, after peer review. To
access the final edited and published work see
http://pubsdc3.acs.org/articlesonrequest/AOR-EW6FuIC6wIh6D9qqEeH
Scaling analysis of the screening length in concentrated electrolytes
The interaction between charged objects in an electrolyte solution is a
fundamental question in soft matter physics. It is well-known that the
electrostatic contribution to the interaction energy decays exponentially with
object separation. Recent measurements reveal that, contrary to the
conventional wisdom given by classic Poisson-Boltzmann theory, the decay length
increases with ion concentration for concentrated electrolytes and can be an
order of magnitude larger than the ion diameter in ionic liquids. We derive a
simple scaling theory that explains this anomalous dependence of the decay
length on ion concentration. Our theory successfully collapses the decay
lengths of a wide class of salts onto a single curve. A novel prediction of our
theory is that the decay length increases linearly with the Bjerrum length,
which we experimentally verify by surface force measurements. Moreover, we
quantitatively relate the measured decay length to classic measurements of the
activity coefficient in concentrated electrolytes, thus showing that the
measured decay length is indeed a bulk property of the concentrated electrolyte
as well as contributing a mechanistic insight into empirical activity
coefficients.Comment: To appear in Physical Review Letter
Nucleocytoplasmic transport: a thermodynamic mechanism
The nuclear pore supports molecular communication between cytoplasm and
nucleus in eukaryotic cells. Selective transport of proteins is mediated by
soluble receptors, whose regulation by the small GTPase Ran leads to cargo
accumulation in, or depletion from the nucleus, i.e., nuclear import or nuclear
export. We consider the operation of this transport system by a combined
analytical and experimental approach. Provocative predictions of a simple model
were tested using cell-free nuclei reconstituted in Xenopus egg extract, a
system well suited to quantitative studies. We found that accumulation capacity
is limited, so that introduction of one import cargo leads to egress of
another. Clearly, the pore per se does not determine transport directionality.
Moreover, different cargo reach a similar ratio of nuclear to cytoplasmic
concentration in steady-state. The model shows that this ratio should in fact
be independent of the receptor-cargo affinity, though kinetics may be strongly
influenced. Numerical conservation of the system components highlights a
conflict between the observations and the popular concept of transport cycles.
We suggest that chemical partitioning provides a framework to understand the
capacity to generate concentration gradients by equilibration of the
receptor-cargo intermediary.Comment: in press at HFSP Journal, vol 3 16 text pages, 1 table, 4 figures,
plus Supplementary Material include
A physical model describing the interaction of nuclear transport receptors with FG nucleoporin domain assemblies
The permeability barrier of nuclear pore complexes (NPCs) controls bulk nucleocytoplasmic exchange. It consists of nucleoporin domains rich in phenylalanine-glycine motifs (FG domains). As a bottom-up nanoscale model for the permeability barrier, we have used planar films produced with three different end-grafted FG domains, and quantitatively analyzed the binding of two different nuclear transport receptors (NTRs), NTF2 and Importin b, together with the concomitant film thickness changes. NTR binding caused only moderate changes in film thickness; the binding isotherms showed negative cooperativity and could all be mapped onto a single master curve. This universal NTR binding behavior âa key element for the transport selectivity of the NPC âwas quantitatively reproduced by a physical model that treats FG domains as regular, flexible polymers, and NTRs as spherical colloids with a homogeneous surface, ignoring the detailed arrangement of interaction sites along FG domains and on the NTR surface
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