University of Heidelberg

ZMBH - Central Services - Core Facility for Mass Spectrometry & Proteomics


The ZMBH Core Facility for Mass Spectrometry and Proteomics provides state-of-the-art services for identification and characterization of proteins to support research of the ZMBH investigators. Capacity permitting, samples from externs can be analyzed at a highly competitive prize.

Dr. Thomas Ruppert
Head of the Mass Spectrometry Department

06221 - 54 6895
(fax: 06221 - 54 5891)

t.ruppert@zmbh.uni-heidelberg.de





Services

  • Protein identification by MALDI-TOF/TOF mass spectrometry
  • Peptide sequencing by nanoESI QTOF mass spectrometry
  • Determination of the exact molecular weight of proteins
  • Multidimensional protein identification technology (MudPIT)

In addition the Core Facility is involved in the development of new methods for analyzing proteins, such as:

  • Identification of posttranslational modifications
  • Determination of the N-terminus and C-terminus of proteins
  • Analysis of protein conformation by amide hydrogen-1H/2H-exchange

It is equipped with modern, high end-instruments for protein and pepitde analytics:

  • Sample preparation: Pipetting robot: Digest Pro (Intavis), nanoHPLC: ULTIMATE (LC-Packings/Dionex), FAMOS and SWITCHOS, Eksigent 1D plus (Eksigent), nanoAcquity (Waters)
  • Mass Spectrometers: Maldi-Tof MS: ULTRAFLEX TOF/TOF (Bruker), ESI-Qtof MS: QSTAR PULSAR (Applied Biosysystems), ESI QTrap MS (ABSciex), ESI-LTQ Orbitrap MS (Thermo)

Participation in ABRF research studies

sPRG 2010 (Determination of phosphorylation sites):
Study letter
Results

sPRG 2010 (qualitative protein analysis)
Study letter
Results are submitted to ABRF

Downloads

Protocols

Protein staining

  • Colloidal Coomassie (EMBL)
  • Blue silver (Candiano et al.)
  • Silver staining (EMBL, should only be used if colloidal coomassie is not sensitive enough for picking the protein spot/band. Silver stained gel plugs must be destained before storage to avoid loss of peptides, Richard S. et al.)

Sample preparation

Links

Data search

Companies

Armin Bosserhoff
06221 - 54 6813



Margrit Ellis
06221 - 54 6896



Hartmut Krischke
06221 - 54 6813



Sebastian Link
06221 - 54 6892



Sabine Merker
06221 - 54 6813


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Publications

Kemmler, S. Stach, M., Knapp, M., Ortiz, J. Pfannstiel, J., Ruppert, T., and Lechner, J. (2009). Mimicking Ndc80 phosphorylation triggers spindel assembly checkpoint signaling. EMBO J 28, 1099-1110

Tegha-Dungu, J., Neumann, B., Reber, S., Krause, R., Erfle, H., Walter, T., Held, M., Rogers, P., Hupfeld, K., Ruppert, T. Ellenberg, J. and Gruss, O. (2008). EML 3 is a nuclear microtubule-binding protein required for the correct alignment of chromosomes in metaphase. J Cell Sci 121, 1718-1726
Supplementary Data

Major T., von Janowsky, B., Ruppert, T., Mogk, A. and Voos, W. (2006). Proteomic analysis of mitochondrial protein turnover: identification of novel substrate proteins of the matrix protease pim 1. Mol Cell Biol 26, 762-776

Colansante, C., Ellis, M., Ruppert, T., and Voncken, F. (2006). Comparative proteomics of glycosomes from bloodstream form and procyclic culture form Trypanosoma burcei brucei. Proteomics 6, 3275-3293

Herrmann, R. and Ruppert, T. (2006). Proteome of Mycoplasma pneumoniae. Methods Biochem Anal 49, 39-56

Kupfahl C., Heinekamp, T., Geginat, G., Ruppert, T., Hartl, A., Hof, H., and Brakhage, A.A. (2006). Deletion of the gliP gene of Aspergillus fumigatus results in loss of gliotoxin production but has no effect on virulence of the fungus in a low-dose mouse infection model. Mol Microbiol.

Gotthardt, D., Blancheteau V., Bosserhoff, A., Ruppert, T., Delorenzi, M., and Soldati, T. (2006). Mol Cell Proteomics

Lee, B., Bauer, H., Melchers, J., Ruppert, T., Rattray, L., Yardley, V., Davioud-Charvet, E., Krauth-Siegel, R.L. (2005). Irreversible inactivation of trypanothione reductase by unsaturated Mannich bases: a divinyl ketone as key intermediate. J Med Chem 48, 7400-7410

Catrein, I. Herrmann, R., Bosserhoff, A., and Ruppert T. (2005). Experimental proof for a signal petpidase I like activity in Mycoplasma pneumoniae, but absence of a gene encoding a conserved bacterial type I SPase, FEBS J 272, 2892-2900

Stubs, D., Fuchs, T.M., Scheider, B., Bosserhoff, A., and Gross, R. (2005). Identification and regulation of cold-inducible factors of Bordetella bronchiseptica. Microbiol 151, 1895-1909

Seifert, U., Liermann, H., Racanelli, V., Halenius, A., Wiese, M., Wedemeyer, H., Ruppert, T., Rispeter, K., Henklein, P., Sijts, A., et al. (2004). Hepatitis C virus mutation affects proteasomal epitope processing. J Clin Invest 114, 250-259.

Estevez, A.M., Lehner, B., Sanderson, C. M., Ruppert, T., and Clayton C. (2003). The roles of intersubunit interactions in exosome stability. J Biol Chem 278, 34943-34951

Schwend, T., Redwanz, I., Ruppert, T., Szenthe, A., and Wink, M. (2003). Analysis of proteins in the spent culture medium of Lupinus albus by electrospray ionisation tandem mass spectrometry. J Chromatogr A 1009, 105-110.

Simons, A., Ruppert, T., Schmidt, C., Pipkorn, R., Reed, J., Masters, C. L., White, A., Cappai, R., Beyreuther, K., Bayer, T., and Multhaup, G. (2002). Evidence for a copper-binding superfamily of the amyloid precursor protein. Biochemistry 41, 9310 - 9320.

Spory, A., Bosserhoff, A., von Rhein, C., Goebel, W., and Ludwig, A. (2002). Differential regulation of multiple proteins of Escherichia coli and Salmonella enterica serovar Typhimurium by the transcriptional regulator SlyA. J Bacteriol 184, 3549-3559.

Papaioannou, M., Ruppert, T., Dotzlaw, H., Dressel, U., and Baniahmad, A. (2002). Protein-protein cross-linking in the use of the eukaryotic eGST-fusion system. Protein Expr Purif 26, 462-466.

Kuckelkorn U, Ruppert T, Strehl B, Jungblut PR, Zimny-Arndt U, Lamer S, Prinz I, Drung I, Kloetzel PM, Kaufmann SH, Steinhoff U. J. Exp. Med. 2002 Apr; 195(8): 983-90

Opitz C, Christina M, Reiss M, Ruppert T, Crisanti A, Soldati D. Intramembrane cleavage of microneme proteins at the surface of the apicomplexan parasite Toxoplasma gondii. EMBO J. 2002 21(7) 1577-1585

Bubeck A, Reusch U, Wagner M, Ruppert T, Muranyi W, Kloetzel PM, Koszinowski UH. The glycoprotein gp48 of murine Cytomegalovirus: Proteasome-dependent cytosolic dislocation and degradation. J Biol Chem. 2001 Nov 13

Stanislawski T, Voss RH, Lotz C, Sadovnikova E, Willemsen RA, Kuball J, Ruppert T, Bolhuis RL, Melief CJ, Huber C, Stauss HJ, Theobald M. Circumventing tolerance to a human MDM2-derived tumor antigen by TCR gene transfer. Nat Immunol. 2001 Oct;2(10):962-70

Knuehl C, Spee P, Ruppert T, Kuckelkorn U, Henklein P, Neefjes J, Kloetzel PM. The murine cytomegalovirus pp89 immunodominant H-2Ld epitope is generated and translocated into the endoplasmic reticulum as an 11-mer precursor peptide. J Immunol. 2001 Aug 1;167(3):1515-21.

Dahlmann B, Ruppert T, Kloetzel PM, Kuehn L. Subtypes of 20S proteasomes from skeletal muscle. Biochimie. 2001 Mar-Apr;83(3-4):295-9.

Dahlmann B, Ruppert T, Kuehn L, Merforth S, Kloetzel PM. Different proteasome subtypes in a single tissue exhibit different enzymatic properties. J Mol Biol. 2000 Nov 10;303(5):643-53.

Hengel H, Reusch U, Geginat G, Holtappels R, Ruppert T, Hellebrand E, Koszinowski UH. Macrophages escape inhibition of major histocompatibility complex class I-dependent antigen presentation by cytomegalovirus. J Virol. 2000 Sep;74(17):7861-8.

Sijts AJ, Standera S, Toes RE, Ruppert T, Beekman NJ, van Veelen PA, Ossendorp FA, Melief CJ, Kloetzel PM. MHC class I antigen processing of an adenovirus CTL epitope is linked to the levels of immunoproteasomes in infected cells. J Immunol. 2000 May 1;164(9):4500-6.

Sijts AJ, Ruppert T, Rehermann B, Schmidt M, Koszinowski U, Kloetzel PM. Efficient generation of a hepatitis B virus cytotoxic T lymphocyte epitope requires the structural features of immunoproteasomes. J Exp Med. 2000 Feb 7;191(3):503-14.

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