TY - JOUR A1 - Xie, J. A1 - Techritz, S. A1 - Haebel, Sophie A1 - Horn, A. A1 - Neitzel, H. A1 - Klose, J. A1 - Schuelke, M. T1 - A two-dimensional electrophoretic map of human mitochondrial proteins from immortalized lymphoblastoid cell lines: a prerequisite to study mitochondrial disorders in patients N2 - Mitochondrial diseases may be caused by numerous mutations that alter proteins of the respiratory chain and of other metabolic pathways in the mitochondrium. For clinicians this disease group poses a considerable diagnostic challenge due to ambiguous genotype-phenotype relationships. Until now, only 30 % of the mitochondriopathies can be diagnosed at the molecular level. We therefore need a new diagnostic tool that offers a wide view on the mitochondrial proteins. Here, we present a method to generate a high-resolution, large-gel two-dimensional gel electrophoretic (2-DE) map of a purified fraction of mitochondrial proteins from Epstein-Barr virus-immortalized lymphoblastoid cell line (LCL). LCLs can be easily obtained from patients and control subjects in a routine clinical setting. They often express the biochemical phenotype and can be cultured to high cell numbers, sufficient to gain enough purified material for 2- DE. In total we identified 166 mitochondrial proteins. Thirteen proteins were earlier not known to be of mitochondrial origin. Thirty-nine proteins were associated with human diseases ranging from respiratory chain enzyme deficiencies to disorders of P-oxidation and amino acid metabolism. This 2-DE map is intended to be the first step to diagnose mitochondrial diseases at the proteomic level Y1 - 2005 SN - 1615-9853 ER - TY - JOUR A1 - Tsukamoto, Junko A1 - Haebel, Sophie A1 - Valenca, Gustavo P. A1 - Peter, Martin G. A1 - FRanco, Telma T. T1 - Enzymatic direct synthesis of acrylic acid esters of mono- and disaccharides N2 - Background: There is an increased need to replace materials derived from fossil sources by renewables. Sugar- cane derived carbohydrates are very abundant in Brazil and are the cheapest sugars available in the market, with more than 400 million tons of sugarcane processed in the year 2007. The objective of this work was to study the prepn. of sugar acrylates from free sugars and free acrylic acid, thus avoiding the previous prepn. of protected sugar derivs., such as glycosides, or activated acrylates, such as vinyl acrylate. Results: Lipase catalyzed esterification of three mono- and two disaccharides with acrylic acid, in the presence or absence of mol. sieves was investigated. The reactions were monitored by high-performance liq. chromatog. (HPLC) and the products were analyzed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. The main products are mono- and diacrylates, while higher esters are formed as minor products. The highest conversion to sugar acrylates was obsd. for the D-glucose and D- fructose, followed by D-xylose and D-maltose. Mol. sieves had no pronounced effect on the conversion. Conclusions: A feasible method is described to produce and to characterize sugar acrylates, including those contg. more than two acrylate groups. The process for prodn. of these higher esters could potentially be optimized further to produce mols. for crosslinking in acrylate polymn. and other applications. The direct enzymic esterification of free carbohydrates with acrylic acid is unprecedented. [on SciFinder (R)]. Y1 - 2008 UR - http://www3.interscience.wiley.com/journal/117946200/grouphome/home.html SN - 0268-2575 ER - TY - JOUR A1 - Stöckigt, Detlef A1 - Haebel, Sophie T1 - Identification of CID fragments from a protonated glutathione conjugate by isotope-specific MS3 experiments in an ion trap Y1 - 1998 ER - TY - JOUR A1 - Strehmel, Veronika A1 - Sarker, A. M. A1 - Lahti, P. M. A1 - Karasz, F. E. A1 - Heydenreich, Matthias A1 - Wetzel, Hendrik A1 - Haebel, Sophie A1 - Strehmel, Bernd T1 - One- and two-photon photochemistry and photophysics of poly(arylenevinylene)s containing a biphenyl moiety N2 - Photochemical and photophysical properties were investigated for poly(arylenevinylene)s containing a flexible biphenyl "hinge" unit by applying one-photon (OP) and two-photon (TP) excitation to explore excited-state properties. The poly(arylenevinylene)s were poly[(2,5-dihexyloxy-p-phenylenevinylene)-alt-(4,4'-dihexyloxy-3,3'-biph enylenevinylene)] (1), poly[(2,5-dihexyloxy-p-phenylenevinylene)-alt-(2,2'-dihexyloxy-3,3'-biph enylenevinylene)] (2), and poly[(2,5-dihexyloxy-p-phenylenevinylene)-alt-(2,2'-biphenylene-vinylene )] (3). Effective emission quantum yields and related photonic properties were evaluated on a realistic per-chromophore basis using effective conjugation lengths based on the Strickler-Berg relationship. intramolecular photocyclization was deduced to occur in the one case where the biphenyl molecular connectivity permitted the reaction, based on matrix- assisted loser desorption/ionization time-of-flight (MALDI-TOF), heteronuclear multiple-quantum coherence (HMQC)-NMR, and gel-permeation chromatography (GPC) results. The various photoprocesses could be induced by either OP or TP excitation, though the first excited singlet state is the photoactive state. The higher excitation energy 1 of the TP excited state favors indirect population of the S, state by electronic coupling between the TP and OP excited states [lambda(max)(TPE) (nm): 726; delta (GM)([9]): 1 = 229, 2 = 215, 3 = 109). Photochemical processes occurring from the lowest OP excited state (S-1) could therefore also be indirectly induced by TP excitation Y1 - 2005 SN - 1439-4235 ER - TY - JOUR A1 - Stalz, Holger A1 - Roth, Udo A1 - Schleuder, Detlev A1 - Macht, Marcus A1 - Haebel, Sophie A1 - Strupat, Kerstin A1 - Peter-Katalinic, Jasna A1 - Hanisch, Franz-Georg T1 - The Geodia cydonium galectin exhibits prototype and chimera-type characteristics and a unique sequence polymorphism within its carbohydrate recognition domain N2 - The ancestral galectin from the sponge Geodia cydonium (GCG) is classified on a structural basis to the prototype subfamily, whereas its carbohydrate-binding specificity is related to that of the mammalian chimera-type galectin-3. This dual coordination reveals GCG as a potential precursor of the later evolved galectin subfamilies, which is reflected in the primary structure of the protein. This study provides evidence that GCG is the LECT1 gene product, while neither a previously described LECT2 gene nor a functional LECT2 gene product was found in the specimen under investigation. The electrophoretically separated protein isomers with apparent molecular masses of 13, 15, and 16 kDa correspond to variants of the LECT1 protein-exhibiting peptide sequence polymorphisms that concern critical positions of the carbohydrate recognition domain (13 kDa: Leu51, Asn55, His130, Gly137; 15 kDa: Ser51, Asn55, Asn130, Gly137; 16 kDa: Ser51, Tyr55, Asn130, Glu137). Four residues, highly conserved in the galectin family, are substituted. None of the residues claimed to be involved in interactions with GalNAc alpha 1-3 moieties at an extended binding subsite of galectin-3 was identified in the corresponding positions of GCG. Apparently, the substitutions do not confer distinct binding characteristics to the GCG variants as evidenced by binding studies with a recombinantly expressed 15-kDa isoform. The natural isoforms as well as the recombinant 15-kDa isoform oligomerize by the formation of non-covalent heteromeric or homomeric complexes. A phosphorylation of the galectin was confirmed neither by mass spectrometry nor by alkaline phosphatase treatment combined with isoelectric focusing Y1 - 2006 UR - http://glycob.oxfordjournals.org/content/16/5/402.full U6 - https://doi.org/10.1093/glycob/cwj086 ER - TY - JOUR A1 - Schweigert, Florian J. A1 - Raila, Jens A1 - Haebel, Sophie T1 - Vitamin A excreted in the urine of canines is associated with a Tamm-Horsfall-like Glycoprotein Y1 - 1998 ER - TY - JOUR A1 - Ritte, Gerhard A1 - Scharf, Anke A1 - Eckermann, Nora A1 - Haebel, Sophie A1 - Steup, Martin T1 - Phosphorylation of transitory starch is increased during degradation N2 - The starch excess phenotype of Arabidopsis mutants defective in the starch phosphorylating enzyme glucan, water dikinase (EC 2.7.9.4) indicates that phosphorylation of starch is required for its degradation. However, the underlying mechanism has not yet been elucidated. In this study, two in vivo systems have been established that allow the analysis of phosphorylation of transitory starch during both biosynthesis in the light and degradation in darkness. First, a photoautotrophic culture of the unicellular green alga Chlamydomonas reinhardtii was used to monitor the incorporation of exogenously supplied P-32 orthophosphate into starch. Illuminated cells incorporated P-32 into starch with a constant rate during 2 h. By contrast, starch phosphorylation in darkened cells exceeded that in illuminated cells within the first 30 min, but subsequently phosphate incorporation declined. Pulse-chase experiments performed with P-32/P-31 orthophosphate revealed a high turnover of the starch-bound phosphate esters in darkened cells but no detectable turnover in illuminated cells. Secondly, leaf starch granules were isolated from potato (Solanum tuberosum) plants grown under controlled conditions and glucan chains from the outer granule layer were released by isoamylase. Phosphorylated chains were purified and analyzed using high performance anion-exchange chromatography and matrix-assisted laser desorption/ionization mass spectrometry. Glucans released from the surface of starch granules that had been isolated from darkened leaves possessed a considerably higher degree of phosphorylation than those prepared from leaves harvested during the light period. Thus, in the unicellular alga as well as in potato leaves, net starch degradation is accompanied with an increased phosphorylation of starch Y1 - 2004 ER - TY - JOUR A1 - Ritte, Gerhard A1 - Heydenreich, Matthias A1 - Mahlow, Sebastian A1 - Haebel, Sophie A1 - Koetting, Oliver A1 - Steup, Martin T1 - Phosphorylation of C6- and C3-positions of glucosyl residues in starch is catalysed by distinct dikinases JF - FEBS letters : the journal for rapid publication of short reports in molecular biosciences N2 - Glucan, water dikinase (GWD) and phosphoglucan, water dikinase (PWD) are required for normal starch metabolism. We analysed starch phosphorylation in Arabidopsis wildtype plants and mutants lacking either GWD or PWD using P-31 NMR. Phosphorylation at both C6- and C3-positions of glucose moieties in starch was drastically decreased in GWD-deficient mutants. In starch from PWD-deficient plants C3-bound phosphate was reduced to levels close to the detection limit. The latter result contrasts with previous reports according to which GWD phosphorylates both C6- and C3-positions. In these studies, phosphorylation had been analysed by HPLC of acid-hydrolysed glucans. We now show that maltose-6-phosphate, a product of incomplete starch hydrolysis, co-eluted with glucose-3-phosphate under the chromatographic conditions applied. Re-examination of the specificity of the dikinases using an improved method demonstrates that C6- and C3-phosphorylation is selectively catalysed by GWD and PWD, respectively. KW - starch phosphorylation KW - GWD KW - PWD KW - P-31 NMR Y1 - 2006 U6 - https://doi.org/10.1016/j.febslet.2006.07.085 SN - 0014-5793 VL - 580 IS - 20 SP - 4872 EP - 4876 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Ritte, Gerhard A1 - Eckermann, Nora A1 - Haebel, Sophie A1 - Lorberth, Ruth A1 - Steup, Martin T1 - Compartmentation of the starch-related R1 protein in higher plants Y1 - 2000 ER - TY - JOUR A1 - Rawel, Harshadrai Manilal A1 - Kroll, Jürgen A1 - Riese-Schneider, Brigitte A1 - Haebel, Sophie T1 - Changes in physico-chemical and enzymatic properties of benzyl isothiocyanate derivatized proteinases Y1 - 1998 ER -