Einfluss des Hitzeschockproteins Hsc70 und des Raf Kinase Inhibitor Proteins RKIP auf den zellulären Transport und die Aktivität des [my]-Opioidrezeptors

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2008

Molecular dynamics of the neuronal Ca 2+ -binding proteins Caldendrin and Calneurons

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2009

Mitochondrial localization ot two brain proteins, p42IP4/centaurin-[alpha]1/ADAP1 and CNP, and their involvement in regulation of mitochondrial Ca 2+

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2010

Modulation of [My]-opioid receptor signal transduction and endocytosis by ADP-ribosylation factor proteins

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2010

Convection and segregation of beads in a flat rotating box

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2012

apoptosis-regulator c-FLIP : functional role in urothelial carcinoma and autoimmunity and identification of novel CD95 DISC-interacting proteins

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2013

Interactions of the adaptor proteins AP2 and 14-3-3 with the presynaptic scaffolding protein Bassoon

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2015

Untersuchung des humanen Proteins NF110 als Bindungspartner viraler RNAs

In der vorliegenden Masterarbeit wurde das Protein nuclear factor 110 (NF110) in vitro rückgefaltet, chromatographisch gereinigt, durch den Zusatz von L-Arginin stabilisiert und seine Affinität gegenüber RNAs untersucht. Das Ausgangsmaterial wurde mittels Dialyse zurückgefaltet und über drei aufeinander folgende Chromatographieschritte gereinigt: mittels Kationenaustauschchromatographie (capture) zur Entfernung von Fremdproteinen, Größenausschlusschromatographie (intermediate) zur Trennung des monomeren Proteins von höher-oligomeren Spezies und der Affinitätschromatographie (polishing). Diese Proteinlösung wurde unter Zusatz von 500 mM L-Arginin gelagert um eine Proteinaggregation zu unterbinden. Durch Streulichtmessungen und eine analytische Ultrazentrifugation konnte erfolgreich nachgewiesen werden, dass NF110 erst in Puffern mit mindestens 200 mM L-Arginin stabil vorliegt. Im Rahmen der vorliegenden Arbeit konnte erfolgreich bestimmt werden, dass NF110 zwischen doppelsträngiger RNA und DNA diskriminiert, während es gegenüber einzel- bzw. doppelsträngiger RNA ähnlich affin ist. Dies stellt einen großen Unterschied zu NF90 dar. Diese C-terminal verkürzte Variante interagiert mit einzelsträngiger RNA in sehr geringerem Umfang als mit doppelsträngiger RNA (Schmidt, 2015). Daher kann vermutet werden, dass der C-Terminus, insbesondere das dort befindliche GQSY-Motiv, für die Spezifität gegenüber den RNAs verantwortlich ist. Ein weiterer Fokus der Arbeit lag auf dem Einfluss von L-Arginin bei der RNA-Bindung. Mit Hilfe einer linearen freien Enthalpiebeziehung (LFER) konnte so eine Dissoziationskonstante für nicht aggregierendes NF110 ohne L-Arginin ermittelt werden. Außerdem wurde ersichtlich, dass dieser Stabilisator zwar die Aggregation hemmt, aber nur marginal die RNA-Bindung des Proteins beeinflusst

Patterns of snake evolution suggested by their proteins : a contribution in celebration of the distinguished scholarship of Robert F. Inger on the occasion of his sixty-fifth birthday / Herbert C. Dessauer, John E. Cadle, Robin Lawson.

n.s. no.34(1987)

Lactacidemia: métodos de dosagem do ácido lático

A critical study of three methods for the determination of lactic acid (EDWARDS, MENDEL & GOLDSCHEIDER, MILLER & MUNTZ) is presented and some modifications are proposed. It was shown t hat more accurate results could be obtained with Edward's technic when an Iena glass filter is connected with the absorption tube. Before the dropping of the permanganate solution it is necessary to pass a current of air through the reaction flask to avoid the oxidation of the non-lactic acid substances which interfere with the reaction. The absorption tube must be maintained at 18°C during the destillation and the titration of the bisulphite binding aldehyde at 4°C. When the sample contains more than 5 mg it is useful to work with greater quantities of the bisulphite. More permanganate is consumed when the lactic acid concentration is higher. The sensivity of the method permits the titration of 0.04 mg to 5 mg of lactic acid in the sample. The calculated error of the method gave 0.018 % and the normal values for blood determined in 20 human cases averaged 10.30 mg per 100 ml (Table VI). MENDEL and GOLDSCHEIDER'S method was modified in the following details: Somogyis deproteinization was performed instead metaphosphoric acid as in the original method; to avoid the evaporation of the acetic aldehyde during the heating time with sulfuric acid a special glass stopped tube is proposed (Fig. 2). The reaction with sulfuric acid and veratrol is performed in an ice bath. Blood proteins precipitants were tried and Somogyi's lattest tecnic showed better results (Table V). Colorimetric readings were done in the PULFRICH photometer using filter S 53 and a 10 mm cup. The method is accurate within an error of 0.23 % and samples of 5 to 70 microg. could be easily determined. Normal values for human blood averaged 10.78 mg per 100 ml. More accurate results were obtained with the technic of MILLER & MUNTZ. Slight modifications were introduced: deproteinization with copper sulfate and sodium tungstate; satured p-hydroxydiphenyl solution according to KOENEMANN which is stable for 5 months when stored in the ice-box. Using the PULFRICH step-photometer the error is 0.17% with samples varying from 0.1 to 10 microg. of lactic acid. The filter employed was S 57 with the 5 mm cup. The method was adapted to 0.1 ml of blood. Normal values for human blood gave an average of 10.58 mg per 100 ml.

Aplicació de l'ACV a l'envasat del vi : "Bag in Box" front a l'envàs de vidre tradicional

L’objectiu d’aquest projecte és la comparació, des del punt de vista ambiental, de l’envasat del vi mitjançant ampolles de vidre i mitjançant el sistema “Bag-in-Box” reutilitzable.

CLOCK-controlled polyphonic regulation of circadian rhythms through canonical and noncanonical E-boxes.

In mammalian circadian clockwork, the CLOCK-BMAL1 complex binds to DNA enhancers of target genes and drives circadian oscillation of transcription. Here we identified 7,978 CLOCK-binding sites in mouse liver by chromatin immunoprecipitation-sequencing (ChIP-Seq), and a newly developed bioinformatics method, motif centrality analysis of ChIP-Seq (MOCCS), revealed a genome-wide distribution of previously unappreciated noncanonical E-boxes targeted by CLOCK. In vitro promoter assays showed that CACGNG, CACGTT, and CATG(T/C)G are functional CLOCK-binding motifs. Furthermore, we extensively revealed rhythmically expressed genes by poly(A)-tailed RNA-Seq and identified 1,629 CLOCK target genes within 11,926 genes expressed in the liver. Our analysis also revealed rhythmically expressed genes that have no apparent CLOCK-binding site, indicating the importance of indirect transcriptional and posttranscriptional regulations. Indirect transcriptional regulation is represented by rhythmic expression of CLOCK-regulated transcription factors, such as Krüppel-like factors (KLFs). Indirect posttranscriptional regulation involves rhythmic microRNAs that were identified by small-RNA-Seq. Collectively, CLOCK-dependent direct transactivation through multiple E-boxes and indirect regulations polyphonically orchestrate dynamic circadian outputs.

Immunocytochemical localisation of microtubule-associated proteins 1b and 2 in the developing rat spinal cord.

The straightforward anatomical organisation of the developing and mature rat spinal cord was used to determine and interpret the time of appearance and expression patterns of microtubule-associated proteins (MAP) 1b and 2. Immunoblots revealed the presence of MAP1b and 2 in the early embryonic rat spinal cord and confirmed the specificity of the used anti-MAP mouse monoclonal antibodies. The immunocytochemical data demonstrated a rostral-to-caudal and ventral-to-dorsal gradient in the expression of MAP1b/2 within the developing spinal cord. In the matrix layer, MAP1b was found in a distinct radial pattern distributed between the membrana limitans interna and externa between embryonal day (E)12 and E15. Immunostaining for vimentin revealed that this MAP1b pattern was morphologically and topographically different from the radial glial pattern which was present in the matrix layer between E13 and E19. The ventral-to-dorsal developmental gradient of the MAP1b staining in the spinal cord matrix layer indicates a close involvement of MAP1b either in the organisation of the microtubules in the cytoplasmatic extensions of the proliferating neuroblasts or neuroblast mitosis. MAP2 could not be detected in the developing matrix layer. In the mantle and marginal layer, MAP1b was abundantly present between E12 and postnatal day (P)0. After birth, the staining intensity for MAP1b gradually decreased in both layers towards a faint appearance at maturity. The distribution patterns suggest an involvement of MAP1b in the maturation of the motor neurons, the contralaterally and ipsilaterally projecting axons and the ascending and descending long axons of the rat spinal cord. MAP2 was present in the spinal cord grey matter between E12 and maturity, which reflects a role for MAP2 in the development as well as in the maintenance of microtubules. The present description of the expression patterns of MAP1b and 2 in the developing spinal cord suggests important roles of the two proteins in various morphogenetic events. The findings may serve as the basis for future studies on the function of MAP1b and 2 in the development of the central nervous system.

Bioluminescent sensor proteins for point-of-care therapeutic drug monitoring.

For many drugs, finding the balance between efficacy and toxicity requires monitoring their concentrations in the patient's blood. Quantifying drug levels at the bedside or at home would have advantages in terms of therapeutic outcome and convenience, but current techniques require the setting of a diagnostic laboratory. We have developed semisynthetic bioluminescent sensors that permit precise measurements of drug concentrations in patient samples by spotting minimal volumes on paper and recording the signal using a simple point-and-shoot camera. Our sensors have a modular design consisting of a protein-based and a synthetic part and can be engineered to selectively recognize a wide range of drugs, including immunosuppressants, antiepileptics, anticancer agents and antiarrhythmics. This low-cost point-of-care method could make therapies safer, increase the convenience of doctors and patients and make therapeutic drug monitoring available in regions with poor infrastructure.