Project Z2

Quantiative methods for the analysis of the spatial and temporal dynamic changes of the proteome during host-pathogen interaction

Project Description

Z2

In project Z2 a sophisticated proteome analysis platform will be provided based on 2D-gel electrophoresis (2D-GE), MALDI-TOF and LC-MS/MS techniques to specific A and Cprojects. 2D-GE is an established technique with high-resolution capacity, which still provides some advantages over LC-MS-based proteomic technologies: information about protein abundance is immediately available and posttranslational modification can readily be located in 2D-gels. For the detection of proteins either classical post-staining methods will be applied or proteins will be labelled prior to electrophoresis with spectrally resolvable fluorescent dyes (2D-differential gel electrophoresis (DIGE)). Additionally, gel-free methods, like LC-MS-based separations are ideal for the comprehensive investigation of hydrophobic and membrane proteins. For such analyses, an analytical nano-LCs and three different types of mass analysers are available, i.e. an ion-trap, a MALDI-TOF/TOF and a Q-TOF device

Besides the temporal also the spatial analyses of changes of the proteome during the interaction of pathogen and host will be studied in collaboration. The secretion of fungal secondary metabolites as well as extracellular proteins will be investigated in situ. For this purpose, MALDI-imaging will be applied which can determine the spatial distribution of unknown compounds (secondary metabolites, peptides) in complex tissues with a single label-free measurement while maintaining the cellular integrity within the tissue.

All data from imaging and protein identification will be transferred to the INF project and will be stored in a freely accessible data warehouse for the integrated storage and visualisation of genome and experimental transcriptome and proteome data of human-pathogenic fungi (data ware house OmniFung).

Principal Investigators

Dr. Olaf Kniemeyer
Dr. Olaf Kniemeyer

Department of Molecular and Applied Microbiology

Leibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute  

olaf.kniemeyer@leibniz-hki.de

PD Dr. Ferdinand von Eggeling
PD Dr. Ferdinand von Eggeling

Institute of Physical Chemistry

Friedrich Schiller University Jena

ferdinand.von_eggeling@med.uni-jena.de

Publications

Author Year Title Journal Links
Hillmann F, Linde J, Beckmann N, Cyrulies M, Strassburger M, Heinekamp T, Haas H, Guthke R, Kniemeyer O, Brakhage AA 2014 The novel globin protein fungoglobin is involved in low oxygen adaptation of Aspergillus fumigatus. Mol Microbiol 93: 539-53 PubMed
Lapp K, Vödisch M, Kroll K, Strassburger M, Kniemeyer O, Heinekamp T, Brakhage AA 2014 Characterisation of the Aspergillus fumigatus detoxification systems for reactive nitrogen intermediates and impact on virulence. Front Microbiol 5: 469 PubMed
Baldin C, Valiante V, Krüger T, Schafferer L, Haas H, Kniemeyer O, Brakhage AA 2015 Comparative proteomics of a tor inducible Aspergillus fumigatus mutant reveals involvement of the tor kinase in iron regulation. Proteomics 15: 2230-43 PubMed
Heinekamp T, Schmidt H, Lapp K, Pähtz V, Shopova I, Köster-Eiserfunke N, Krüger T, Kniemeyer O, Brakhage AA 2014 Interference of Aspergillus fumigatus with the immune response. Semin Immunopathol 37: 141-52 PubMed

Altwasser R, Baldin C, Weber J, Guthke R, Kniemeyer O, Brakhage AA, Linde J, Valiante V

2015

Network modeling reveals cross talk of MAP kinases during adaptation to caspofungin stress in Aspergillus fumigatus.

PLoS One 10: e0136932 PubMed
Teutschbein J, Simon S, Lother J, Springer J, Hortschansky P, Morton CO, Löffler J, Einsele H, Conneally E, Rogers TR, Guthke R, Brakhage AA, Kniemeyer O 2016 Proteomic profiling of serological responses to Aspergillus fumigatus antigens in patients with invasive aspergillosis.

J Proteome Res 15: 1580-91

PubMed
Moyes DL, Wilson D, Richardson JP, Mogavero S, Tang SX, Wernecke J, Höfs S, Gratacap RL, Robbins J, Runglall M, Murciano C, Blagojevic M, Thavaraj S, Förster TM, Hebecker B, Kasper L, Vizcay G, Iancu SI, Kichik N, Häder A, Kurzai O, Luo T, Krüger T, Kniemeyer O, Cota E, Bader O, Wheeler RT, Gutsmann T, Hube B, Naglik JR 2016

Candidalysin is a fungal peptide toxin critical for mucosal infection.

Nature 532: 64-8 PubMed
Bacher P, Heinrich F, Stervbo U, Nienen M, Vahldieck M, Iwert C, Vogt K, Kollet J, Babel N, Sawitzki B, Schwarz C, Bereswill S, Heimesaat MM, Heine G, Gadermaier G, Asam C, Assenmacher M, Kniemeyer O, Brakhage AA, Ferreira F, Wallner M, Worm M, Scheffold A 2016 Regulatory T cell specificity directs tolerance versus allergy against aeroantigens in humans. Cell 167: 1067-78.e16 PubMed
Krüger T, Luo T, Schmidt H, Shopova I, Kniemeyer O 2015 Challenges and strategies for proteome analysis of the interaction of human pathogenic fungi with host immune cells. Proteomes 3: 467-95 Proteomes
Heinekamp T, Schmidt H, Lapp K, Pähtz V, Shopova I, Köster-Eiserfunke N, Krüger T, Kniemeyer O, Brakhage AA 2015 Interference of Aspergillus fumigatus with the immune response. Semin Immunopathol 37: 141-52 PubMed
Luo T, Krüger T, Knüpfer U, Kasper L, Wielsch N, Hube B, Kortgen A, Bauer M, Giamarellos-Bourboulis EJ, Dimopoulos G, Brakhage AA, Kniemeyer O 2016 Immunoproteomic analysis of antibody responses to extracellular proteins of Candida albicans revealing the importance of glycosylation for antigen recognition. J Proteome Res 15: 2394-406 PubMed
Kniemeyer O, Ebel F, Krüger T, Bacher P, Scheffold A, Luo T, Strassburger M, Brakhage AA 2016 Immunoproteomics of Aspergillus for the development of biomarkers and immunotherapies. Proteomics Clin Appl 10: 910-21 PubMed
Pohlers S, Martin R, Krüger T, Hellwig D, Hänel F, Kniemeyer O, Saluz HP, Van Dijck P, Ernst JF, Brakhage A, Mühlschlegel FA, Kurzai O 2017 Lipid Signaling via Pkh1/2 regulates fungal CO2 sensing through the kinase Sch9. mBio 8 pii: e02211-16 PubMed