Signal peptide peptidases and gamma-secretase: Cousins of the same protease family?

Signal peptide peptidase (SPIP) is an unusual aspartyl protease, which mediates clearance of signal peptides by proteolysis within the endoplasmic reticulum (ER). Like presenilins, which provide the proteolytically active subunit of the,gamma-secretase complex, SPP contains a conserved GxGD motif in its C-terminal domain which is critical for its activity. While SPIP is known to be an aspartyl protease of the GxGD type, several presenilin homologues/SPP-like proteins (PSHs/ SPPL) of unknown function have been identified by database searches. In contrast to SPP and SPPL3, which are both restricted to the endoplasmic reticulum, SPPL2b is targeted through the secretory pathway to endosomes/lysosomes. As suggested by the differential subcellular localization of SPPL2b and SPPL3 distinct phenotypes were found upon antisense gripNA-mediated knockdown in zebrafish. spp and sppl3 knockdowns in zebrafish result in cell death within the central nervous system, whereas reduction of sppl2b expression causes erythrocyte accumulation in an enlarged caudal vein. Moreover, expression of D/A mutants of the putative C-terminal active sites of spp, sppl2, and spp13 produced phenocopies of the respective knockdown phenotypes. These data suggest that all investigated PSHs/SPPLs are members of the novel family of GxGD aspartyl proteases. More recently, it was shown that SPPL2b utilizes multiple intramembrane cleavages to liberate the TNF(x intracellular domain into the cytosol and to release the C-terminal counterpart into the lumen. These findings suggest common principles of intramembrane proteolysis by GxGD type aspartyl proteases. In this article,we will review the similarities of SPPs and gamma-secretase based on recent findings by us and others

Quasi-free Compton Scattering and the Polarizabilities of the Neutron

Differential cross sections for quasi-free Compton scattering from the proton and neutron bound in the deuteron have been measured using the Glasgow/Mainz tagging spectrometer at the Mainz MAMI accelerator together with the Mainz 48 cm $\oslash$ $\times$ 64 cm NaI(Tl) photon detector and the G\"ottingen SENECA recoil detector. The data cover photon energies ranging from 200 MeV to 400 MeV at $\theta^{LAB}_\gamma=136.2^\circ$. Liquid deuterium and hydrogen targets allowed direct comparison of free and quasi-free scattering from the proton. The neutron detection efficiency of the SENECA detector was measured via the reaction $p(\gamma,\pi^+ n)$. The "free" proton Compton scattering cross sections extracted from the bound proton data are in reasonable agreement with those for the free proton which gives confidence in the method to extract the differential cross section for free scattering from quasi-free data. Differential cross sections on the free neutron have been extracted and the difference of the electromagnetic polarizabilities of the neutron have been obtained to be $\alpha-\beta= 9.8\pm 3.6(stat){}^{2.1}_1.1(syst)\pm 2.2(model)$ in units $10^{-4}fm^3$. In combination with the polarizability sum $\alpha +\beta=15.2\pm 0.5$ deduced from photoabsorption data, the neutron electric and magnetic polarizabilities, $\alpha_n=12.5\pm 1.8(stat){}^{+1.1}_{-0.6}\pm 1.1(model)$ and $\beta_n=2.7\mp 1.8(stat){}^{+0.6}_{-1.1}(syst)\mp 1.1(model)$ are obtained. The backward spin polarizability of the neutron was determined to be $\gamma^{(n)}_\pi=(58.6\pm 4.0)\times 10^{-4}fm^4$

Neutron polarizabilities investigated by quasi-free Compton scattering from the deuteron

Measuring Compton scattered photons and recoil neutrons in coincidence, quasi-free Compton scattering by the neutron has been investigated at MAMI (Mainz) at $theta^{lab}_\gamma=136^o$ in an energy range from 200 to 400 MeV. From the data a polarizability difference of $\alpha_n - \beta_n = 9.8 \pm 3.6(stat)^{+2.1}_{-1.1}(syst)\pm 2.2(model)$ in units of $10^{-4}fm^3$ has been determined. In combination with the polarizability sum $\alpha_n+\beta_n= 15.2\pm 0.5$ deduced from photo absorption data, the neutron electric and magnetic polarizabilities, $\alpha_n=12.5\pm 1.8(stat)^{+1.1}_{-0.6}(syst)\pm 1.1(model)$ and $\beta_n = 2.7\mp 1.8(stat)^{+0.6}_{-1.1}(syst)\mp 1.1(model)$, are obtained

Quasi-free π0\pi^0 Photoproduction from the Bound Nucleon

Differential cross-sections for quasi-free $\pi^0$ photoproduction from the proton and neutron bound in the deuteron have been measured for $E_\gamma= 200 - 400$ MeV at $\theta^{\rm lab}_\gamma = 136.2^\circ$ usind the Glasgow photon tagger at MAMI, the Mainz 48 cm $\varnothing$ $\times$ 64 cm NaI(Tl) photon detector and the G\"ottingen SENECA recoil detector. For the proton measurements made with both liquid deuterium and liquid hydrogen targets allow direct comparison of "free" $\pi^0$ photoproduction cross-sections as extracted from the bound proton data with experimental free cross sections which are found to be in reasonable agreement below 320 MeV. At higher energies the "free" cross sections extracted from quasifree data are significantly smaller than the experimental free cross sections and theoretical predictions based on multipole analysis. For the first time, "free" neutron cross sections have been extracted in the $\Delta$-region. They are also in agreement with the predictions from multipole analysis up to 320 MeV and significantly smaller at higher photon energies

The helicity amplitudes A1/2_{1/2} and A3/2_{3/2} for the D13(1520)_{13}(1520) resonance obtained from the γ⃗p⃗→pπ0\vec{\gamma} \vec{p} \to p \pi^0 reaction}

The helicity dependence of the $\vec{\gamma} \vec{p} \to p \pi^0$ reaction has been measured for the first time in the photon energy range from 550 to 790 MeV. The experiment, performed at the Mainz microtron MAMI, used a 4$\pi$-detector system, a circularly polarized, tagged photon beam, and a longitudinally polarized frozen-spin target. These data are predominantly sensitive to the $D_{13}(1520)$ resonance and are used to determine its parameters.Comment: 5 pages, 4 figure

First measurement of the Gerasimov-Drell-Hearn integral for Hydrogen from 200 to 800 MeV

A direct measurement of the helicity dependence of the total photoabsorption cross section on the proton was carried out at MAMI (Mainz) in the energy range 200 < E_gamma < 800 MeV. The experiment used a 4$\pi$ detection system, a circularly polarized tagged photon beam and a frozen spin target. The contributions to the Gerasimov-Drell-Hearn sum rule and to the forward spin polarizability $\gamma_0$ determined from the data are 226 \pm 5 (stat)\pm 12(sys) \mu b and -187 \pm 8 (stat)\pm 10(sys)10^{-6} fm^4, respectively, for 200 < E_\gamma < 800 MeV.Comment: 6 pages, 3 figures, 3 table

Acer rubrum Wats.

https://thekeep.eiu.edu/herbarium_specimens_byname/21742/thumbnail.jp

Expression and Characterization of Drosophila Signal Peptide Peptidase-Like (sppL), a Gene That Encodes an Intramembrane Protease

Intramembrane proteases of the Signal Peptide Peptidase (SPP) family play important roles in developmental, metabolic and signaling pathways. Although vertebrates have one SPP and four SPP-like (SPPL) genes, we found that insect genomes encode one Spp and one SppL. Characterization of the Drosophila sppL gene revealed that the predicted SppL protein is a highly conserved structural homolog of the vertebrate SPPL3 proteases, with a predicted nine-transmembrane topology, an active site containing aspartyl residues within a transmembrane region, and a carboxy-terminal PAL domain. SppL protein localized to both the Golgi and ER. Whereas spp is an essential gene that is required during early larval stages and whereas spp loss-of-function reduced the unfolded protein response (UPR), sppL loss of function had no apparent phenotype. This was unexpected given that genetic knockdown phenotypes in other organisms suggested significant roles for Spp-related proteases

Helicity dependence of the γ→p→→nπ+π0 reaction in the second resonance region

The helicity dependence of the total cross section for the reaction has been measured for the first time at incident photon energies from 400 to 800 MeV. The measurement was performed with the large acceptance detector DAPHNE at the tagged photon beam facility of the MAMI accelerator in Mainz. This channel is found to be excited predominantly when the photon and proton have a parallel spin orientation, due to the intermediate production of the D13 resonance. peerReviewe

Alzheimer's Disease-Linked Mutations in Presenilin-1 Result in a Drastic Loss of Activity in Purified γ-Secretase Complexes

BACKGROUND: Mutations linked to early onset, familial forms of Alzheimer's disease (FAD) are found most frequently in PSEN1, the gene encoding presenilin-1 (PS1). Together with nicastrin (NCT), anterior pharynx-defective protein 1 (APH1), and presenilin enhancer 2 (PEN2), the catalytic subunit PS1 constitutes the core of the γ-secretase complex and contributes to the proteolysis of the amyloid precursor protein (APP) into amyloid-beta (Aβ) peptides. Although there is a growing consensus that FAD-linked PS1 mutations affect Aβ production by enhancing the Aβ1-42/Aβ1-40 ratio, it remains unclear whether and how they affect the generation of APP intracellular domain (AICD). Moreover, controversy exists as to how PS1 mutations exert their effects in different experimental systems, by either increasing Aβ1-42 production, decreasing Aβ1-40 production, or both. Because it could be explained by the heterogeneity in the composition of γ-secretase, we purified to homogeneity complexes made of human NCT, APH1aL, PEN2, and the pathogenic PS1 mutants L166P, ΔE9, or P436Q. METHODOLOGY/PRINCIPAL FINDINGS: We took advantage of a mouse embryonic fibroblast cell line lacking PS1 and PS2 to generate different stable cell lines overexpressing human γ-secretase complexes with different FAD-linked PS1 mutations. A multi-step affinity purification procedure was used to isolate semi-purified or highly purified γ-secretase complexes. The functional characterization of these complexes revealed that all PS1 FAD-linked mutations caused a loss of γ-secretase activity phenotype, in terms of Aβ1-40, Aβ1-42 and APP intracellular domain productions in vitro. CONCLUSION/SIGNIFICANCE: Our data support the view that PS1 mutations lead to a strong γ-secretase loss-of-function phenotype and an increased Aβ1-42/Aβ1-40 ratio, two mechanisms that are potentially involved in the pathogenesis of Alzheimer's disease