TY - JOUR A1 - Bahrke, Sven A1 - Einarsson, Jon M. A1 - Gislason, Johannes A1 - Haebel, Sophie A1 - Peter-Katalinic, Jasna T1 - Characterization of chitooligosaccharides by mass spectrometry N2 - Heterochitooligosaccharides of DP 6, DP 9, and DP 12 were evaluated using established methods of derivatization and matrix-assisted laser desorption ionization post source decay mass spectrometry. Y1 - 2003 SN - 82-471-5901-5 ER - TY - THES A1 - Bahrke, Sven A1 - Einarsson, Jon M. A1 - Gislason, Johannes A1 - Haebel, Sophie A1 - Peter-Katalinic, Jasna A1 - Peter, Martin G. T1 - Characterization of chitooligosaccharides by mass spectrometry Y1 - 2003 SN - 82-47-15901-5 ER - TY - JOUR A1 - Cederkvist, F. Henning A1 - Zamfir, Alina D. A1 - Bahrke, Sven A1 - Eijsink, Vincent G. H. A1 - Sorlie, Morten A1 - Peter-Katalinic, Jasna A1 - Peter, Martin G. T1 - Identification of a high-affinity-binding oligosaccharide by (+) nanoelectrospray quadrupole time-of-flight tandem mass spectrometry of a noncovalent enzyme-ligand complex Y1 - 2006 UR - http://www3.interscience.wiley.com/cgi-bin/jhome/26737/ U6 - https://doi.org/10.1002/anie.200503168 SN - 1433-7851 ER - TY - THES A1 - Bahrke, Sven T1 - Mass spectrometric analysis of chitooligosaccharides and their interaction with proteins T1 - Massenspektrometrische Analyse von Chitooligosacchariden und ihre Wechselwirkungen mit Proteinen N2 - Chitooligosaccharides are composed of glycosamin and N-acetylglycisamin residues. Gel permeations chromatography is employed for the separation of oligomers, cation exchange chromatography is used for the separation of homologes and isomers. Trideuterioacetylation of the chitooligosaccharides followed by MALDI-TOF mass spectrometry allowes for the quantitation of mixtures of homologes. vMALDI LTQ multiple-stage MS is employed for quantitative sequencing of complex mixtures of heterochitooligosaccharides. Pure homologes and isomers are applied to biological assays. Chitooligosaccahrides form high-affinity non-covalent complexes with HC gp-39 (human cartilage glycoprotein of 39 kDa). The affinity of the chitooligosaccharides depends on DP, FA and the sequence of glycosamin and N-acetylglycosamin moieties. (+)nanoESI Q TOF MS/MS is used for identification of a high-affinity binding chitooligosaccharide of a non-covalent chitinase B - chitooligosaccharide complex. DADAA is identified as the heterochitoisomer binding with highest affinity and biostability to HC gp-39. Fluorescence based enzyme assays confirm the results. N2 - Chitooligosaccharide sind aus Glycosamin und N-Acetylglycosamun aufgebaut. Gelpermeationschromatographie wird für die Trennung von Oligomeren verwendet, die Kationenaustauschchromatographie wird zur Trennung von homologen- und Isomerengemischen angewendet. Trideuterioacetylierung der Chitooligosaccharide gefolgt von einer Analyse mittels MALDI-TOF MS erlaubt die quantitative Analyse von Homologengemischen. vMALDI LTQ multiple-stage MS wird angewendet zur Sequenzanalyse und Quantifizierung komplexer Gemische von Heterochitooligosacchariden. Reine Homologe und Isomere werden für biologische Assays verwendet. Dabei zeigt sich, dass Chitooligosaccharide mit HC gp-39 hochaffine Komplexe bilden. Die Affinität der Chitooligosaccharide hängt vom DP, FA und der Sequenz der Chitooligosaccharide ab. (+)nanoESI Q TOF MS/MS wird erfolgreich angewendet zur Identifizierung eines Chitooligosaccharides, das mit hoher Affinität an Chitinase B (Serratia marcescens) bindet. DADAA wurde als die Sequenz des Isomers identifiziert, das mit höchster Affinität und Biostabilität an aktive Chitinase B bindet. Fluoreszenz basierte Enzymassays konnten dieses Ergebnis bestätigen. KW - Chitooligosaccharide KW - HPLC KW - Massenspektrometrie KW - Chitolektine KW - Chitinase KW - Chitooligosaccharides KW - HPLC KW - Mass Spectrometry KW - Chitolectins KW - Chitinase Y1 - 2008 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-20179 ER - TY - JOUR A1 - Einarsson, Jon M. A1 - Bahrke, Sven A1 - Sigurdsson, Bjarni Thor A1 - Ng, Chuen-How A1 - Petersen, Petur Henry A1 - Sigurjonsson, Olafur E. A1 - Jonsson, Halldor A1 - Gislason, Johannes A1 - Thormodsson, Finnbogi R. A1 - Peter, Martin G. T1 - Partially acetylated chitooligosaccharides bind to YKL-40 and stimulate growth of human osteoarthritic chondrocytes JF - Biochemical and biophysical research communications N2 - Recent evidences indicating that cellular kinase signaling cascades are triggered by oligomers of N-acetylglucosamine (ChOS) and that condrocytes of human osteoarthritic cartilage secrete the inflammation associated chitolectin YKL-40, prompted us to study the binding affinity of partially acetylated ChOS to YKL-40 and their effect on primary chondrocytes in culture. Extensive chitinase digestion and filtration of partially deacetylated chitin yielded a mixture of ChOS (Oligomin(TM)) and further ultrafiltration produced T-ChOS(TM), with substantially smaller fraction of the smallest sugars. YKL-40 binding affinity was determined for the different sized homologues, revealing micromolar affinities of the larger homologues to YKL-40. The response of osteoarthritic chondrocytes to Oligomin(TM) and T-ChOS(TM) was determined, revealing 2- to 3-fold increases in cell number. About 500 mu g/ml was needed for Oligomin(TM) and around five times lower concentration for T-ChOS(TM), higher concentrations abolished this effect for both products. Addition of chitotriose inhibited cellular responses mediated by larger oligosaccharides. These results, and the fact that the partially acetylated T-ChOS(TM) homologues should resist hydrolysis, point towards a new therapeutic concept for treating inflammatory joint diseases. KW - Cell culture KW - Chitolectin KW - Chitooligosaccharides KW - Chondrocytes KW - High affinity binding KW - Rheumatoid arthritis KW - YKL-40 Y1 - 2013 U6 - https://doi.org/10.1016/j.bbrc.2013.02.122 SN - 0006-291X VL - 434 IS - 2 SP - 298 EP - 304 PB - Elsevier CY - San Diego ER - TY - JOUR A1 - Haebel, Sophie A1 - Bahrke, Sven A1 - Peter, Martin G. T1 - Quantitative sequencing of complex mixtures of heterochitooligosaccharides by vMALDI-linear ion trap mass spectrometry N2 - Heterochitooligosaccharides possess interesting biol. properties. Isobaric mixts. of such linear heterochitooligosaccharides can be obtained by chem. or enzymic degrdn. of chitosan. However, the sepn. of such mixts. is a challenging anal. problem which is so far unresolved. It is shown that these isobaric mixts. can be sequenced and quantified simultaneously using std. derivatization and multistage tandem mass spectrometric techniques. A linear ion trap mass spectrometer equipped with a vacuum matrix-assisted laser desorption ionization (vMALDI) source is used to perform MS2 as well as MS3 expts. [on SciFinder (R)]. Y1 - 2007 UR - http://pubs.acs.org/loi/ancham U6 - https://doi.org/10.1021/Ac062254u SN - 0003-2700 ER - TY - JOUR A1 - Bahrke, Sven A1 - Einarsson, Jon M. A1 - Gislason, Johannes A1 - Haebel, Sophie A1 - Letzel, Matthias C. A1 - Peter-Katalinic, Jasna A1 - Peter, Martin G. T1 - Sequence analysis of chitooligosaccharides by matrix-assisted laser desorption ionization postsource decay mass spectrometry N2 - Oligosaccharides composed of 2-acetamido-2-deoxy-D-glucopyranose (GlcNAc) and/or 2-amino-2-deoxy-D- glucopyranose (GlcN) were prepd. by chem. degrdn. of chitin or chitosan and sepd. by gel permeation chromatog. Oligosaccharides obtained after enzymic hydrolysis of chitosan [FA 0.19] with a fungal chitinase were derivatized by reductive amination with 2-aminoacridone and sequenced by matrix-assisted laser desorption ionization time-of-flight postsource decay (PSD) mass spectrometry (MS). The sequence of a trimer, D1A2, was established as D-A-A. The compn. of a hexamer D3A3 was .apprx.65% D-A-D-D-A-A and 35% D-D-A-D-A-A. The PSD MS of a nonamer D5A4-amac revealed four isobaric species D-X-Y-D-X-Y-D-A-A, where A is GlcNAc, D is GlcN, and X and Y (X ¹ Y) are mutually either D or A. This structure motif was also obsd. in a dodecamer D7A5 which was composed of eight isobaric sequences of the general formula (D-X-Y)3- D-A-A. Y1 - 2002 ER -