Rab11 is a small GTP-binding protein that in cultured mammalian cells has been shown to be concentrated in the pericentriolar endosomal recycling compartment and to play a key role in passage of the recycling transferrin receptor through that compartment [Ullrich, O., Reinsch, S., Urbé, S. To obtain insights into the site(s) of action of rab11 within the recycling pathway, we have now compared the effects on recycling at 37°C of overexpression of wild-type rab11 and various mutant forms of this protein in cells that had been loaded with transferrin at either 37°C or 16°C. We show that incubation at 16°C blocks passage of endocytosed transferrin into the recycling compartment and that, whereas the rab11 dominant negative mutant form (S25N) inhibits transferrin recycling after interiorization at either temperature, the wildtype rab11 and constitutively active mutant (Q70L) have no inhibitory effect on the recycling of molecules that were interiorized at 16°C. This differential inhibitory effect shows that two distinct pathways for recycling are followed by the bulk of the transferrin molecules interiorized at the two different temperatures. The incapacity of the constitutively active form of rab11 (Q70L) to inhibit recycling of molecules interiorized at 16°C is consistent with their recycling taking place directly from sorting endosomes, in a process that does not require hydrolysis of GTP on rab11. The fact that the dominant negative (S25N) form of rab11 inhibits recycling of molecules interiorized at both temperatures indicates that activation of rab11 by GTP is required for exit of transferrin from sorting endosomes, regardless of whether this exit is toward the recycling compartment or directly to the plasma membrane.

Plasmonic metal nanostructures have shown great potential in sensing, photovoltaics, imaging and biomedicine, principally due to the enhancement of local electric field by light-excited surface plasmons, i.e., collective oscillation of conduction band electrons. Thin films of nanoporous gold have received a great deal of interest due to the unique 3-dimensional bicontinuous nanostructures with high specific surface area. However, in the form of semi-infinite thin films, nanoporous gold exhibits weak plasmonic extinction and little tunability in the plasmon resonance, because the pore size is much smaller than the wavelength of light. Here we show that by making nanoporous gold in the form of disks of sub-wavelength diameter and sub-100 nm thickness, these limitations can be overcome. Nanoporous gold disks not only possess large specific surface area but also high-density, internal plasmonic "hot-spots" with impressive electric field enhancement, which greatly promotes plasmon-matter interactions as evidenced by spectral shifts in the surface plasmon resonance. In addition, the plasmonic resonance of nanoporous gold disks can be easily tuned from 900 to 1850 nm by changing the disk diameter from 300 to 700 nm. Furthermore, nanoporous gold disks can be fabricated as either bound on a surface or as non-aggregating colloidal suspension with high stability.

No author has a financial or proprietary interest in any material or method mentioned.

Nanoporous gold disks (NPGDs) with 400 nm diameter, 75 nm thickness, and 13 nm pores exhibit large specific surface area and effective photothermal light harvesting capability with a conversion efficiency of 56%. A potential application is demonstrated by light-gated, multi-step molecular release of the pre-adsorbed R6G fluorescent dye on arrayed NPGDs.

We report that 3 nm diameter CdTe quantum dots can generate enhanced Raman scattering. The Raman signal of 4-mercaptopyridine (4-Mpy) adsorbed on CdTe quantum dots shows a 104 enhancement compared with that of bulk 4-Mpy. The enhanced phenomenon based on CdTe quantum dots also provides new insight to help understand the enhancement mechanism. A charge-transfer mechanism is most likely responsible for the observed enhancement, since plasmon resonances are ruled out. This study points to the possibility of using quantum dots, with chemisorption in some important practical systems, such as the application of quantum dots as nanosized building blocks and markers in biological imaging.

Rab8 is a small GTP-binding protein that plays a role in vesicular transport from the trans-Golgi network to the basolateral plasma membrane in polarized epithelial cells (MDCK), and to the dendritic surface in hippocampal neurons. As is the case for most other rab proteins, the precise molecular interactions by which rab8 carries out its function remain to be elucidated. Here we report the identification and the complete cDNA-derived amino acid sequence of a murine rab8-interacting protein (rab8ip) that specifically interacts with rab8 in a GTP-dependent manner.Rab8ip displays 93% identity with the GC kinase, a serine/ threonine protein kinase recently identified in human lymphoid tissue that is activated in the stress response. Like the GC kinase, rab8ip has protein kinase activity manifested by autophosphorylation and phosphorylation of the classical serine/threonine protein kinase substrates, myelin basic protein and casein. When coexpressed in transfected 293T cells, rab8 and the rab8ip/GC kinase formed a complex that could be recovered by immunoprecipitation with antibodies to rab8. Cell fractionation and immunofluorescence analyses indicate that in MDCK cells endogenous rab8ip is present both in the cytosol and as a peripheral membrane protein concentrated in the Golgi region and basolateral plasma membrane domains, sites where rab8 itself is also located. In light of recent evidence that rab proteins may act by promoting the stabilization of SNARE complexes, the specific GTP-dependent association of rab8 with the rab8ip/GC kinase raises the possibility that rab-regulated protein phosphorylation is important for vesicle targeting or fusion. Moreover, the rab8ip/GC kinase may serve to modulate secretion in response to stress stimuli.

We have identified and cloned the cDNA for a 912-aa protein, rab11BP, that interacts with the GTPcontaining active form of rab11, a GTP-binding protein that plays a critical role in receptor recycling. Although rab11BP is primarily cytosolic, a significant fraction colocalizes with rab11 in endosomal membranes of both the sorting and recycling subcompartments. In vitro binding of rab11 to native rab11BP requires partial denaturation of the latter to expose an internal binding site located between residues 334 and 504 that is apparently masked by the C-terminal portion of the protein, which includes six repeats known as WD40 domains. Within the cell, rab11BP must undergo a conformational change in which the rab11-binding site becomes exposed, because when coexpressed with rab11 in transfected cells the two proteins formed abundant complexes in association with membranes. Furthermore, although overexpression of rab11BP did not affect transferrin recycling, overexpression of a truncated form of the protein, rab11BP(1-504), that includes the rab11-binding site but lacks the WD40 domains inhibited recycling as strongly as does a dominant negative rab11 mutant protein that does not bind GTP. Strikingly, the inhibition caused by the truncated rab11BP was prevented completely when the cells also expressed a C-terminally deleted, nonprenylatable form of rab11 that, by itself, has no effect on recycling. We propose that rab11BP is an effector for rab11, whose association with this GTP-binding protein is dependent on the action of another membrane-associated factor that promotes the unmasking of the rab11-binding site in rab11BP.

Understanding interactions between Nafion (perfluorosulfonic acid) and Pt catalysts is important for the development and deployment of proton exchange membrane fuel cells. However, study of such interactions is challenging and Nafion/Pt interfacial structure remains elusive. In this study, adsorption of Nafion ionomer on Au and Pt surfaces was investigated for the first time by in situ surface-enhanced Raman spectroscopy. The study is made possible by the use of uniform SiO(2)@Au core-shell particle arrays which provides very strong enhancement of Raman scattering. The high surface sensitivity offered by this approach yields insightful information on interfacial Nafion structure. Through spectral comparison of several model compounds, vibration assignments of SERS bands were made. The SER spectra suggest the direct interaction of sulfonate group with the metal surfaces, in accord with cyclic voltammetric results. Comparison of present SERS results with previous IR spectra was briefly made.