Showing papers by "Friedrich Lottspeich published in 2014"

Protein signatures of oxidative stress response in a patient specific cell line model for autism

Abstract: Known genetic variants can account for 10% to 20% of all cases with autism spectrum disorders (ASD). Overlapping cellular pathomechanisms common to neurons of the central nervous system (CNS) and in tissues of peripheral organs, such as immune dysregulation, oxidative stress and dysfunctions in mitochondrial and protein synthesis metabolism, were suggested to support the wide spectrum of ASD on unifying disease phenotype. Here, we studied in patient-derived lymphoblastoid cell lines (LCLs) how an ASD-specific mutation in ribosomal protein RPL10 (RPL10[H213Q]) generates a distinct protein signature. We compared the RPL10[H213Q] expression pattern to expression patterns derived from unrelated ASD patients without RPL10[H213Q] mutation. In addition, a yeast rpl10 deficiency model served in a proof-of-principle study to test for alterations in protein patterns in response to oxidative stress. Protein extracts of LCLs from patients, relatives and controls, as well as diploid yeast cells hemizygous for rpl10, were subjected to two-dimensional gel electrophoresis and differentially regulated spots were identified by mass spectrometry. Subsequently, Gene Ontology database (GO)-term enrichment and network analysis was performed to map the identified proteins into cellular pathways. The protein signature generated by RPL10[H213Q] is a functionally related subset of the ASD-specific protein signature, sharing redox-sensitive elements in energy-, protein- and redox-metabolism. In yeast, rpl10 deficiency generates a specific protein signature, harboring components of pathways identified in both the RPL10[H213Q] subjects’ and the ASD patients’ set. Importantly, the rpl10 deficiency signature is a subset of the signature resulting from response of wild-type yeast to oxidative stress. Redox-sensitive protein signatures mapping into cellular pathways with pathophysiology in ASD have been identified in both LCLs carrying the ASD-specific mutation RPL10[H213Q] and LCLs from ASD patients without this mutation. At pathway levels, this redox-sensitive protein signature has also been identified in a yeast rpl10 deficiency and an oxidative stress model. These observations point to a common molecular pathomechanism in ASD, characterized in our study by dysregulation of redox balance. Importantly, this can be triggered by the known ASD-RPL10[H213Q] mutation or by yet unknown mutations of the ASD cohort that act upstream of RPL10 in differential expression of redox-sensitive proteins.

Ex vivo processing for maturation of Arabidopsis KDEL-tailed cysteine endopeptidase 2 (AtCEP2) pro-enzyme and its storage in endoplasmic reticulum derived organelles

Abstract: Ricinosomes are specialized ER-derived organelles that store the inactive pro-forms of KDEL-tailed cysteine endopeptidases (KDEL-CysEP) associated with programmed cell death (PCD). The Arabidopsis genome encodes three KDEL-CysEP (AtCEP1, AtCEP2, and AtCEP3) that are differentially expressed in vegetative and generative tissues undergoing PCD. These Arabidopsis proteases have not been characterized at a biochemical level, nor have they been localized intracellularly. In this study, we characterized AtCEP2. A 3xHA-mCherry-AtCEP2 gene fusion including pro-peptide and KDEL targeting sequences expressed under control of the endogenous promoter enabled us to isolate AtCEP2 “ex vivo”. The purified protein was shown to be activated in a pH-dependent manner. After activation, however, protease activity was pH-independent. Analysis of substrate specificity showed that AtCEP2 accepts proline near the cleavage site, which is a rare feature specific for KDEL-CysEPs. mCherry-AtCEP2 was detected in the epidermal layers of leaves, hypocotyls and roots; in the root, it was predominantly found in the elongation zone and root cap. Co-localization with an ER membrane marker showed that mCherry-AtCEP2 was stored in two different types of ER-derived organelles: 10 μm long spindle shaped organelles as well as round vesicles with a diameter of approximately 1 μm. The long organelles appear to be ER bodies, which are found specifically in Brassicacae. The round vesicles strongly resemble the ricinosomes first described in castor bean. This study provides a first evidence for the existence of ricinosomes in Arabidopsis, and may open up new avenues of research in the field of PCD and developmental tissue remodeling. Electronic supplementary material The online version of this article (doi:10.1007/s11103-013-0157-6) contains supplementary material, which is available to authorized users.

Evaluation of determination methods of the Si/Al contact resistance of screen-printed passivated emitter and rear solar cells

Abstract: The Si/Al contact resistance of narrow p-type contact areas of crystalline silicon solar cells is a crucial parameter for the further development and improvement of passivated emitter and rear (PERC) solar cells. In this paper, two methods for determining the rear contact resistance are analyzed and improved: the series resistance analysis and the transfer length method (TLM). We use an experimental set of PERC solar cells with varying metallization fraction, and we analyze the experiment using numerical device simulation. In the first method, the resistive losses are extracted from the I-V-curves of the PERC cells, and the rear contact resistance is separated by subtracting the analytically calculated base contribution. The state-of-the-art analytical calculations deviate significantly from our numerical simulations, causing a high level of uncertainty that we do not recommend this method (an upper limit of the rear contact resistance per line contact is determined to be about 2 Ω cm). In the second method, the rear line contacts are separated and measured with the TLM. Still, the series resistance loss in the base has to be quantified considering a thick base and the rear line contact geometry. We separate the base component by numerical device simulations and find that the rear contact resistance per line contact is below 0.5 Ω cm. This implies that rear contact resistance reduces cell efficiency of screen-printed PERC solar cells only by about 0.1%abs for metallization fractions higher than 3.5%, i.e., the rear contact resistance is currently no significant loss mechanism in our PERC solar cells. All of our improved evaluations lead to lower resistance values than previously reported and with a reduced uncertainty. The TLM method is suggested to be favorable.

DJ-1 protein and Apolipoprotein E are new targets for LVVYPW in diabetic rat brain. Dopaminergic system in the striatum of healthy rats is also target for hemorphin

Abstract: hemorphin, calcineurin, DJ-1 protein. ApolipoE, dopamine A growing body of data demonstrates that hemorphins, a family of endogenous nonclassical opioid peptides derived from hemoglobin (Hb) [22], exhibit a wide spectrum of biological activity by affecting different receptor’s function (e.g. µ-, δ- and к-opioid receptors [39], angiotensin (Ang) IV receptor (AT