Research

 

The goals of our research are to elucidate protein-protein interactions as well as interactions between proteins and small bioactive ligands with the aim for biomedical and pharmaceutical application. Methods employed comprise chemical cross-linking, gel electrophoresis, multi-dimensional liquid chromatography, bioinformatics, and mass spectrometry, including the use of an LTQ-Orbitrap XL mass spectrometer (ThermoFisherScientific) and nano-HPLC (high-performance liquid chromatography) / MALDI (matrix-assisted laser desorption / ionization) TOF (time-of-flight) / TOF mass spectrometry (Ultraflex III, Bruker Daltonik). We also develop gel-free approaches for global proteome studies and determine non-covalent interactions by electrospray ionization (ESI) mass spectrometry. Novel analytical methods are developed as they are applied to the present problems.


 

 Publications

Original Papers

  • Fritzsche, R., Ihling, C.H., Götze, M., Sinz, A., Optimizing the enrichment of cross-linked products for mass spectrometric protein analysis, Rapid Commun. Mass Spectrom., in press.

  • Sproß, J., Sinz, A., Monolithic columns with immobilized monomeric avidin:preparation and application for affinity chromatography, Anal.Bioanal. Chem., online available.

  • Sproß, J., Sinz, A., Monolithic media for applications in affinity chromatography, J. Sep. Sci., online available.

  • Krause, K., Boisnard, A., Ihling, C., Ludgate, M., Eszlinger, M., Krohn, K., Sinz, A., Führer, D., Comparative proteomic analysis to dissect differences in signal transductionin activating TSH receptor mutations in the thyroid, Int. J. Biochem. Cell Biol. 44, 290-301 (2012)

  • Götze, M., Pettelkau, J., Schaks, S., Bosse, K., Ihling, C.H., Krauth, F., Fritzsche, R., Kühn, U., Sinz, A., StavroX – A Software for Analyzing Cross-Linked Products in Protein Interaction Studies, J. Am. Soc. Mass Spectrom. 23, 76-87 (2012)

  • Krause, K., Prawitt, S., Eszlinger, M., Ihling, C., Sinz, A., Schierle, K., Gimm, O., Dralle, H., Steinert, F., Sheu, S.Y., Schmid, K.W., Führer, D., Dissecting molecular events in thyroid neoplasia provides evidence for distinct evolutionof follicular thyroid adenoma and carcinoma, Am. J. Pathol. 179, 3066-3074 (2012)

  • Soboh, B., Pinske, C., Kuhns, M., Waclawek, M., Ihling, C., Trchounian, K., Trchounian, A., Sinz, A., Sawers, G., The respiratory molybdo-selenoprotein formate dehydrogenases of Escherichia coli have hydrogen: benzyl viologen oxidoreductase activity, BMC Microbiol. 11, 173 (2011)

  • Pinske, C., Krüger, S., Soboh, B., Ihling, C., Kuhns, M., Braussemann, M., Jaroschinsky, M., Sauer, C., Sargent, F., Sinz, A., Sawers, R.G., Efficient electron transfer from hydrogen to benzyl viologen by the [NiFe]-hydrogenases of Escherichia coli is dependent on the coexpression of the iron-sulfur cluster-containing small subunit, Arch. Microbiol. 193, 893-903 (2011)

  • Kuhfahl, S., Hauburger, A., Thieme,T., Groppe, J., Ihling, C., Tomic, S., Schutkowski, M., Sinz, A.,Schwarz. E., Identification of a core domain within the proregion of bone morphogenetic proteins that interacts with the dimeric, mature domain, Biochem. Biophys. Res. Commun. 408, 300-305 (2011)

  • Nimptsch, A., Schibur, S., Ihling, C., Sinz, A., Riemer, T., Huster, D.,Schiller, J., Quantitative Analysis of Dentaured Collagen by Collagenase Digestion and subsequent MALDI-TOF Mass Spectrometry, Cell Tissue Res. 343, 605-617 (2011)

  • Müller, M.Q., Zeiser, J.J., Dreiocker, F., Pich, A., Schäfer, M., Sinz, A.,

    A universal matrix-assisted laser desorption/ionization cleavable cross-linker for protein structure analysis, Rapid Commun Mass Spectrom. 25, 155-161 (2011)

  • Kalkhof, S., Witte, K., Ihling, C.H., Müller, M.Q., Keller, M.V., HaehnS., Smyth, N., Paulsson, M., Sinz, A., A Novel Disulfide Patternin Laminin-Type Epidermal Growth Factor-Like (LE) Modules of Laminin ß1 and γ1 Chains, Biochemistry 49, 8359-8366 (2010)
  • Schröder-Tittmann, K., Bosse-Doenecke, E., Reedtz-Runge, S., Ihling,C., Sinz, A., Tittmann, K., Rudolph, R., Recombinant Expression, inVitro Refolding, and Biophysical Characterization of the Human Glucagon-likePeptide-1 Receptor, Biochemistry 49, 7956-65 (2010)
  • Kalkhof, S., Haehn, S., Paulsson, M., Smyth , N., Meiler, J., Sinz, A.,Computational Modeling of Laminin N-TerminalDomains Using Sparse Distance Constraints from Disulfide Bondsand Chemical Cross-Linking, Proteins: Structure, Function andBioinformatics 78, 3409-3427 (2010)
  • Müller, M.Q., Dreiocker, F., Ihling, C.H., Schäfer, M., Sinz, A., A Novel CleavableCross-Linker for ProteinStructure Analysis:Reliable Identification of Cross-Linking Products by TandemMS, Anal. Chem. 82, 6958-6968 (2010)
  • Müller, M.Q., Dreiocker, F., Ihling, C.H., Schäfer, M., Sinz, A.,Fragmentation Behavior of aThiourea-Based Reagent for Protein Structure Analysis by Collision-InducedDissociative Chemical Cross-Linking,J. Mass. Spectrometry 45, 880-891 (2010)
  • Temme. C., Zhang,L., Kremmer, E., Ihling, C., Chartier, A., Sinz,A., Simonelig, M., Wahle, E., Subunitsof the Drosophila CCR4-NOT complex and their roles in mRNA deadenylation, RNA 16, 1356-1370 (2010)
  • Hoffmann, A., Müller, M.Q.,Gloser, M., Sinz, A., Rudolph, R.,Pfeifer, S., Recombinant production of bioactive human TNF-alpha by SUMO-fusionsystem - High yields from shake-flask culture, Protein Expr. Purif. 72, 238-243 (2010)
  • Sproß, J. and Sinz, A., A Capillary Monolithic Trypsin Reactor for Efficient Protein Digestion in Online and Offline Coupling to ESI and MALDI Mass Spectrometry, Anal. Chem. 82, 1434-1443 (2010)
  • Dreiocker, F., Müller., M.Q., Sinz, A.* and Schäfer, M.*, Collision-InducedDissociative Chemical Cross-Linking Reagent for Protein StructureCharacterization: Applied Edman Chemistry in the Gas Phase, J. Mass. Spectrometry 45, 178-189 (2010)
  • Sproß, J. and Sinz, A., Immobilized Monolithic Enzyme Reactors for Application in Proteomics and Pharmaceutics, Anal. Bioanal. Chem. 3951583-1588 (2009)
  • Krauth, F., Ihling, C.H., Rüttinger, H.H. and Sinz,A., Heterobifunctional Isotope-labeled Amine-ReactivePhoto-Cross-Linker for Structural Investigation of Proteins by MALDI-TOF/TOFand ESI-LTQ-Orbitrap Mass Spectrometry, RapidCommun. Mass Spectrom. 23, 2811-2818 (2009)
  • Dimova, K., Kalkhof, S., Pottratz, I., Ihling, C., Rodriguez, F., Liepold, T., Brose, N., Sinz, A.* and Jahn, O.*, Structural Insights into the Calmodulin / Munc13 Interaction by Cross-Linking and Mass Spectrometry, Biochemistry 48, 5908-5921 (2009)
  • Müller, M.Q., de Koning, L. J., Schmidt, A., Ihling, C., Syha, Y., Rau, O., Mechtler, K., Schubert-Zsilavecz, M. and Sinz, A., An Innovative Method to Study Receptor-Drug Interactions by Mass Spectrometry, J. Med. Chem. 52, 2875-2879 (2009)
  • MacDougall, D., Wachten, S., Ciruela, A., Sinz, A. and Cooper, D.M.F., Separate Elements within a Single IQ-like Motif in Adenylyl Cyclase Type 8 Impart Ca2+/CaM Stimulation and Autoinhibition, J. Biol. Chem. 284, 15573-15588 (2009)
  • Manolaridis, I., Mumtsidu, E., Konarev, P., Makhov, A.M., Fullerton, S.F., Sinz, A., Kalkhof, S., McGeehan, J.E., Carey, P.D., Griffith, J.D., Svergun, D. Kneale, G.G. and Tucker, P.A., Structural and Biophysical Characterization of the Components of the Herpes Simplex Virus-1 Replication Origin, J. Biol. Chem. 284, 16343-16353 (2009)
  • Kölbel. K., Ihling, C., Bellmann-Sickert, K., Neundorf, I., Beck-Sickinger, A.G., Sinz, A., Kühn, U. and Wahle, E., T ype I arginine methyl transferases PRMT1 and 3 act distributively, J. Biol. Chem. 284, 8274-8282 (2009)
  • Müller, M.Q., Roth, C., Sträter, N. and Sinz, A., Expression and Purification of the Ligand-Binding Domain of Peroxisome Proliferator-Activated Receptor alpha (PPARalpha), Prot. Exp. Purif. 62, 185-189 (2008), DOI information: 10.1016/j.pep.2008.07.016
  • Kalkhof, S. and Sinz, A., Chances and Pitfalls of Chemical Cross-Linking with Amine-Reactive N-Hydroxy Succinimide Esters, Anal. Bioanal. Chem. 392, 305-312 (2008)
  • Kalkhof, S., Haehn, S., Ihling, C., Smyth, N. and Sinz, A., Determination of Disulfide Bond Patterns in Laminin ß1 Chain N-Terminal Domains by Nano-High Performance Liquid Chromatography/Matrix-Assisted Laser Desorption/Ionization Time-of-Flight/Time-of-Flight Mass Spectrometry, Rapid Commun. Mass Spectrom. 22, 1933–1940 (2008)

 

  • Schulz, D.M., Kalkhof, S., Schmidt, A., Ihling, C., Stingl, C., Mechtler, K., Zschörnig, O. and Sinz, A., Annexin A2 / p11 Interaction: New insights Into Annexin A2 Tetramer Structure by Chemical Cross-Linking, High-Resolution Mass Spectrometry, and Computational Modeling, Proteins 69, 254-269 (2007)
  • Pluder, F., Sinz, A. and Beck-Sickinger, A.G., Proliferative Effect of Calcitonin Gene-Related Peptide is Induced by at least Five Proteins as Identified by Proteome Profiling, Chem. Biol. Drug. Des. 69, 14-22 (2007)
  • Stingl, C., Ihling, C., Ammerer, G., Sinz, A. and Mechtler, K., Application of Different Fragmentation Techniques for the Analysis of Phosphopeptides using a Hybrid Linear Ion Trap-FTICR Mass Spectrometer, Biochim. Biophys. Acta- Proteins and Proteomics 1764, 1842–1852 (2006)
  • Ihling, C., Schmidt, A., Kalkhof, S., Stingl, C., Mechtler, K., Haack, M., Schulz, D.M., Cooper, D.M.F., Beck-Sickinger, A.G. and Sinz, A., Isotope-Labeled Cross-Linkers and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry for Structural Analysis of a Protein / Peptide Complex, J. Am. Soc. Mass Spectrom. 17, 1100-1113 (2006)
  • Schmitt, A., Meiners, I., Schmitt, J., Nöller, J., Ihling, C., Münch, G., Sinz, A. and Nieber, K., Two analytical methods to study the interaction of AGEs with cell surface proteins, J. Biochem. Biophys. Methods 65, 121-136 (2005)
  • Sinz, A., Kalkhof, S. and Ihling, C., Mapping Protein Interfaces by a Trifunctional Cross-Linker Combined with MALDI-TOF and ESI-FTICR Mass Spectrometry, J. Am. Soc. Mass Spectrom. 16, 1921-1931 (2005)
  • Schmidt, A., Kalkhof, S., Ihling, C., Cooper, D.M.F. and Sinz, A., Mapping Protein Interfaces by Chemical Cross-Linking and FTICR Mass Spectrometry: Application to a Calmodulin / Adenylyl Cyclase 8 Peptide Complex, Eur. J. Mass Spectrom. 11, 525-534 (2005)
  • Ihling, C. and Sinz, A., Proteome Analysis of Escherichia coli using High-Performance Liquid Chromatography and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry, Proteomics 5, 2029-2042 (2005)
  • Kalkhof, S., Ihling, C., Mechtler, K. and Sinz, A., Chemical Cross-Linking and High-Performance Fourier Transform Ion Cyclotron Resonance Mass Spectrometry for Protein Interaction Analysis: Application to a Calmodulin / Target Peptide Complex, Anal. Chem. 77, 495-503 (2005)
  • Berger, K., Wissmann, D., Ihling, C., Kalkhof, S., Beck-Sickinger, A., Sinz, A., Paschke, R., Führer, D., Quantitative proteome analysis in benign thyroid nodular disease using the fluorescent ruthenium II tris(bathophenanthroline disulfonate) stain, Mol. Cell. Endocrinol. 227, 21-30 (2004)
  • Sinz, A. and Wang, K., Mapping Spatial Proximities of Sulfhydryl Groups in Proteins Using a Fluorogenic Cross-Linker and Mass Spectrometry, Anal. Biochem. 331, 27-32 (2004)
  • Schulz, D.M., Ihling, C., Clore, G.M. and Sinz, A., Mapping the Topology and Determination of a Low Resolution Three-Dimensional Structure of the Calmodulin-Melittin Complex by Chemical Cross-Linking and High-Resolution FTICR Mass Spectrometry: Direct Demonstration of Multiple Binding Modes, Biochemistry 43, 4703-4715 (2004)
  • Schulz, D.M. and Sinz, A., Chemisches Cross-Linking und Massenspektrometrie zur strukturellen und funktionellen Charakterisierung von Proteinen, BIOSpektrum 10, 45-48 (2004)
  • Sinz, A., Chemical Cross-Linking and Mass Spectrometry for Mapping Three-Dimensional Structures of Proteins and Protein Complexes, J. Mass Spectrom. 38, 1225-1237 (2003)
  • Sinz, A., Jin, A.J. and Zschörnig, O., Evaluation of the Metal Binding Properties of a Histidine-Rich Fusogenic Peptide by Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry, J. Mass Spectrom. 38, 1150-1159 (2003)
  • Dihazi, G.H. and Sinz, A., Mapping Low-Resolution Three-Dimensional Protein Structures Using Chemical Cross-Linking and Fourier Transform Ion-Cyclotron Resonance Mass Spectrometry, Rapid Commun. Mass Spectrom. 17, 2005-2014 (2003)
  • Ihling, C., Berger, K., Höfliger, M.M., Führer, D., Beck-Sickinger, A.G. and Sinz, A., Nano-High-Performance Liquid Chromatography Combined with Nano-Electrospray Ionization Fourier Transform Ion-Cyclotron Resonance Mass Spectrometry for Proteome Analysis, Rapid Commun. Mass Spectrom. 17, 1240-1246 (2003)
  • Sinz, A., Bantscheff, M., Mikkat, S., Ringel, B., Drynda, S., Kekow, J., Thiesen, H.J. and Glocker, M.O., Mass spectrometric proteome analyses of synovial fluids and plasmas from patients suffering from rheumatoid arthritis and comparison to reactive arthritis or osteoarthritis, Electrophoresis 23, 3445-3456 (2002)
  • Sinz, A. and Wang, K., Mapping protein interfaces with a fluorogenic cross-linker and mass spectrometry: Application to nebulin-calmodulin complexes, Biochemistry 40, 7903-7913 (2001)
  • Kawamura, M., Eisenhofer, G., Kopin, I.J., Kador, P.F., Lee, Y.S., Tsai, J.-Y., Fujisawa, S., Lizak, M.J., Sinz, A. and Sato, S., Aldose reductase: A key enzyme in norepinephrine metabolism in rats, Biochem. Pharmacol. 58, 517-524 (1999)
  • Sinz, A., Matusch, R., van Elsaecker, F., Santisuk, T., Chaichana, S. and Reutrakul, V., Phenolic Compounds from Anomianthus dulcis, Phytochemistry 50, 1069-1072 (1999)
  • Sinz, A., Matusch, R., Kaempchen, T., Fiedler, W., Schmidt, J., Santisuk, T., Chaichana, S., Reutrakul, V. and Wangcharoentrakul, S., Novel Acetogenins from the Leaves of Dasymaschalon sootepense, Helv. Chim. Acta 81, 1608-1615 (1998)

  • Sinz, A., Matusch, R., Santisuk, T., Chaichana, S. and Reutrakul, V., Flavonol Glycosides from Dasymaschalon sootepense, Phytochemistry 47, 1393-1396 (1998)

 

Review ArticlesReviews and Book Chapters 

  • Schaks, S., Maucher, D., Ihling, C.H., Sinz, A., Investigation of a Calmodulin / Peptide Complex by Chemical Cross-Linking and High-Resolution Mass Spectrometry, Biomacromolecular Mass Spectrometry, König, S (ed.), Nova Science Publishers, 2012 (ISSN:1936-444X)

     

  • Müller, M.Q. and Sinz, A., Chemical Cross-Linking and High-Resolution Mass Spectrometry to Study Protein-Drug Interactions, M. Bantscheff (ed.) Meth. Mol. Biol. 205-218 (2012)

  • Sinz, A., Investigation of Protein-Protein Interactions in Living Cells by Chemical Cross-Linking and Mass Spectrometry, Anal. Bioanal. Chem. 397, 3433-3440 (2010)

  • Mouradov, D., King, G., Ross, I.L., Forwood, J.K., Hume, D.A., Sinz, A., Martin, J.L., Kobe, B, Huber, T., Protein structure determination using a combination of cross-linking, mass spectrometry, and molecular modeling, Methods Mol Biol. 426, 459-474 (2008)

  • Sinz, A., Investigation of Protein-Ligand Interactions by Mass Spectrometry, ChemMedChem 16, 425-431 (2007)

  • Sinz, A., Isotope-Labeled Photoaffinity Reagents and Mass Spectrometry to Identify Protein-Ligand Interactions (Highlight Artikel), Angew. Chem. Int. Ed. 46, 660–662 (2007); Angew. Chem. 119, 670–673 (2007)

  • Sinz, A., Chemical Cross-Linking and Mass Spectrometry to Map Three-Dimensional Protein Structures and Protein-Protein Interactions, Mass Spectrom. Rev. 25, 663-682 (2006)

  • Ihling, C. and Sinz, A., Fourier Transform Ion Cyclotron Resonance Mass Spectrometry for Protein Structure Analysis, J. Biomacromol. Mass Spectrom. 1, 37-53 (2005)

  • Sinz, A., Chemical Cross-Linking and FTICR Mass Spectrometry for Protein Structure Characterization, Anal. Bioanal. Chem. 381, 44-47 (2005)

  • Sinz, A., Chemical Cross-Linking and Mass Spectrometry for Mapping Three-Dimensional Structures of Proteins and Protein Complexes, J. Mass Spectrom. 38, 1225-1237 (2003) 

  • Ihling, C. and Sinz, A., Fourier Transform Ion Cyclotron Resonance Mass Spectrometry for Protein Structure Analysis, Principles of Biomacromolecular Mass Spectrometry, König, S (ed.), Nova Science Publishers, 2008 (ISBN 1-60021-063-5)

  • Sinz, A., Chemical Cross-Linking for Protein Structure Characterization, Encyclopedia of Mass Spectrometry, vol. 6, 822-830 (2007)

  • Sinz, A., Chemical Cross-Linking and Mass Spectrometry for Investigation of Protein-Protein Interactions, Mass Spectrometry of Protein Interactions, Downard, K.M. (ed.), Wiley-Interscience, 2007 (ISBN 978-0-471-79373-1)

   

Further Reviews 

  • Schaks, S., Kalkhof, S., Krauth, F., Jahn, O. and Sinz, A., Chemisches Cross-Linking und MS zur Untersuchung von Proteinkomplexen, BIOspektrum 17, 524-527 (2011)

  • Sinz, A. and Ceglarek, U., Untersuchung von Proteinen und Metaboliten im klinischen Labor – was können moderne Massenspektrometer leisten? Management & Krankenhaus 7, 19 (2011) 

  • Sproß, J. and Sinz, A., HPLC mit monolithischen Säulen, Deutsche Apotheker Zeitung 30, 46-48 (2009)
  • Krauth, F. and Sinz, A.,  Doping – Arzneimittelmissbrauch der besonderen Art, Pharm. Zeitung 153, 3150-3153 (2008)
  • Sinz, A. and Deininger, S.O., Massenspektrometrie zur Detektion von Biomarkern, Pharm. Unserer Zeit 37, 280-281 (2008)
  • Sinz, A., Die Bedeutung der Mutterkorn-Alkaloide als Arzneistoffe, Pharm. Unserer Zeit 37, 306-309 (2008)
  • Sinz, A., Massenspektrometrie in der Proteomik, Deutsche Apotheker Zeitung 29, 41-45 (2006)
  • Ihling, C. and Sinz, A., FTICR-Massenspektrometrie in der Proteinanalytik, Bioforum 27, 56-57 (2004), Spectrométrie de masse FTICR dans l’analyse des protéines, Bioforum France 3, 30-31 (2004)
  • Schulz, D.M. and Sinz, A., Chemisches Cross-Linking und Massenspektrometrie zur strukturellen und funktionellen Charakterisierung von Proteinen, BIOSpektrum 10, 45-48 (2004)

 


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