Kinetics of the CO oxidation reaction on Pt(111) studied by in situ high-resolution x-ray photoelectron spectroscopy.

High-resolution x-ray photoelectron spectroscopy has been used to study the kinetics of the CO oxidation reaction on a Pt(111) surface in situ. The study focuses on the interaction of a preadsorbed p(2x2) layer of atomic oxygen with CO dosed using a supersonic molecular beam. Measurements of O 1s and C 1s spectra at 120 K show that CO adsorbs on the oxygen precovered substrate, but no reaction occurs. A maximum CO coverage of 0.23 ML (monolayer) is observed, with CO exclusively bound on on-top sites. In accordance with the literature, bridge sites are blocked by the presence of atomic oxygen. The reaction of CO with preadsorbed O to CO(2) is studied isothermally in a temperature range between 275 and 305 K. The reaction rate initially increases with CO pressure, but saturates at 9x10(-7) mbar. The data indicate that a certain amount of disordered oxygen within the p(2x2) layer acts as a starting point of the reaction and for a given temperature reacts with a higher rate than O in the well-ordered oxygen p(2x2) phase. For the reaction of CO with this ordered phase, the results confirm the assumption of a reaction mechanism, which is restricted to the edges of compact oxygen islands. The activation energy of the reaction is determined to (0.53+/-0.04) eV, with a prefactor of 4.7x10(6+/-0.7) s(-1).

Direct visualization of the gamma secretase-generated carboxyl-terminal domain of the amyloid precursor protein: association with Fe65 and translocation to the nucleus.

Amyloid-beta, the peptide that deposits as senile plaques in Alzheimer's disease, is derived from the amyloid precursor protein (APP) by a gamma secretase-mediated intramembranous cleavage. In addition to amyloid-beta, this cleavage produces a carboxyl-terminal intracellular fragment which has an unknown function. The carboxyl-terminal domain of APP interacts in the cytoplasm with an adapter protein, Fe65. We demonstrate by laser scanning confocal microscopy that a gamma secretase generated APP carboxyl-terminal domain, tagged with green fluorescent protein (GFP), translocates to the nucleus in a manner dependent upon stabilization by the adapter protein Fe65; APP which has been mutated to block interactions with Fe65 cannot be detected in the nucleus. The APP-CT domain continues to interact with Fe65 in the nucleus, as determined by both colocalization and fluorescence resonance energy transfer (FRET). Visualization of the APP-CT-Fe65 complex in the nucleus may serve as a readout for processes that modify gamma secretase release of APP-CT.

Demonstration by fluorescence resonance energy transfer of two sites of interaction between the low-density lipoprotein receptor-related protein and the amyloid precursor protein: role of the intracellular adapter protein Fe65.

Amyloid-beta, the major constituent of senile plaques in Alzheimer's disease, is derived from the amyloid precursor protein (APP) by proteolysis. Kunitz protease inhibitor (KPI) containing forms of APP (APP751/770) interact with a multifunctional endocytic receptor, the low-density lipoprotein receptor-related protein (LRP), which modulates its proteolytic processing affecting production of amyloid-beta. We used fluorescence resonance energy transfer (FRET) using labeled LRP and APP in H4 cell line to examine the subcellular localization and the molecular domains involved in the APP-LRP interaction. KPI-containing forms of APP (APP770) demonstrated FRET with LRP that was sensitive to the LRP inhibitor receptor-associated protein (RAP), suggesting an interaction between the extracellular domains of APP770 and LRP. APP695 also interacts with LRP to lesser degree (as measured by extracellular domain probes), and this ectodomain interaction is not altered by RAP. By using C-terminally tagged LRP and APP, we demonstrate a second site of interaction between the C termini of both APP695 and APP770 and the C terminus of LRP, and that the interactions at these regions are not sensitive to RAP. We next examined the possibility that the C-termini APP-LRP interaction was mediated by Fe65, an adaptor protein that interacts with the cytoplasmic tails of LRP and APP. FRET studies confirmed a close proximity between the amino Fe65 phosphotyrosine binding (PTB) domain and LRP cytoplasmic domain and between the carboxyl Fe65 PTB domain and the APP cytoplasmic domain. These findings demonstrate that LRP interaction with APP occurs via both extracellular and intracellular protein interaction domains.