Protein Technologies

Expertise in recombinant protein technologies and biophysical characterization

 

The mission of the Protein Technologies Facility (ProTech) is to further research in molecular and cell biology, protein biochemistry, and structural biology by overcoming major bottlenecks in these fields.

Our core services include molecular cloning, protein production and purification, and biophysical characterization of proteins.

SERVICES

Molecular Cloning

We offer customized generation of DNA constructs for recombinant protein production as a stand-alone service or as the first step in a protein production project. We are specialized in generating multi-gene expression constructs for production of protein complexes.

Recombinant Protein Production and Purification

We offer recombinant protein production in E. coli, Drosophila S2 cells, insect cells using the baculovirus expression vector system (BEVS), and HEK293 cells.

ProTech uses several optimized baculovirus bacmids, licensed from Geneva Biotech. We can also provide fermentation services through our partner Eucodis Bioscience.

Protein purification is offered as a full service at different scales. Due to limited capacity, for projects requiring purification of multiple proteins we also offer training and access to the equipment in our facility.

Biophysical Characterization

We offer protein characterization services using a variety of Biophysical techniques to study protein stability, oligomeric state, and secondary structure, or to analyze biomolecular interactions.

We also provide instrument training and access for some of these methods. To gain an overview of the techniques available, please see the Equipment section. More information can be found on the ProTech Wiki, accessible via MyVBCF. To discuss your biophysical characterization project, please contact Arthur Sedivy.

EQUIPMENT

The NanoTemper Prometheus is a multi-purpose, label-free instrument measuring both intrinsic protein fluorescence and scattering while applying a defined temperature profile. Structural changes (folding/unfolding) as well as aggregation can be monitored over time, as a function of temperature, or under different chemical conditions. Since a sensitive fluorescence detection setup is used, only a minute protein sample is needed (>5µg/ml, 10µl). The Prometheus serves not only as a quality control instrument but also as a tool to find optimum conditions by using buffer screens to optimize stability. Using the Prometheus as a tool for day-to-day protein quality checks will improve the reproducibility of subsequent experiments that use purified proteins for structural and interaction studies while screening for potential binders can open up subsequent avenues of investigation.

The CreoptixTM WAVE is a novel, highly sensitive label-free biosensor used to analyze the affinity and kinetics of molecular interactions. The CreoptixTM WAVE provides similar information to the well-known Surface Plasmon Resonance biosensors but with increased sensitivity and lower noise, a superior “no-clog” microfluidics system allowing the detection and characterization of interactions in crude samples, and the ability to measure a very broad kinetic range (ka=103 – 3x109 M-1 sec-1; kd = 10-5 – 5 sec-1).  Additionally, sample consumption, although dependent on the system studied, is very low. Combined with user friendly and robust analysis software, the CreoptixTM WAVE is ideal to study many different sample types and systems including those previously inaccessible using conventional biosensors.

OmniSEC

The OmniSEC system combines analytical size exclusion chromatography with right angle light scattering and measurement of refractive index, to accurately determine protein molecular weights and concentrations. It can be used to analyze the oligomeric state of proteins and protein complexes and to quantify the different species present in a sample. A full UV-vis spectrum and intrinsic viscosity measurements enable the accurate determination of extinction coefficients and shape/size of protein samples.

PEAQ-ITC

The PEAQ-ITC instrument can be used to perform isothermal titration calorimetry experiments to measure biomolecular interaction parameters. The change in enthalpy upon ligand binding is directly measured by the instrument and can then be used to determine the dissociation constant, stoichiometry, and other thermodynamic parameters of the binding interaction. Interactions in the high nanomolar to high micromolar range can be determined using the PEAQ-ITC.

MASS-1

The MASS-1 instrument is a system for measuring biomolecular interactions by Surface Plasmon Resonance (SPR). In addition to determining dissociation constants, SPR is ideal for measuring kinetics of interactions (kon and koff).

The SPR service is only possible as a full-service. In exceptional cases, access is offered to experienced users.

Aekta Purifier

ProTech has three Aekta Purifier/Aekta PURE systems that can be booked by users after appropriate training, so that purifications can be performed by users in our lab. Additional purification equipment, such as a sonicator and centrifuges, can also be booked after training.

Dynapro II DLS Plate Reader

DLS is a label free technique that analyzes particle size distribution in a bulk sample. It is mainly used for protein quality control, as you can detect size distributions of molecules and therefore aggregation and oligomerization of your protein of interest. The Dynapro II plate reader can also be used for screening or long-term stability measurements. Experiments can be carried out for you; alternatively, you may receive training and can then book the instrument yourself.

Chirascan Plus CD Spectrometer

Circular Dichroism is a spectroscopic technique that uses circularly polarized light to study the structure of chiral molecules, such as proteins. The CD spectrum of a protein can be used to determine protein secondary structure or some aspects of tertiary structure. CD can also be used to study protein stability using thermal melt analysis to determine protein melting temperatures (Tm). We perform CD on a Chirascan Plus CD spectrometer from Applied Photophysics.  Experiments can be carried out for you; alternatively, you may receive training and can then book the instrument yourself.

Monolith NT.115 BLUE/RED

MST is a novel technique used to determine molecular affinities. Thermophoretic changes upon molecular interaction can be monitored by fluorescent labeling or via tryptophan fluorescence, revealing binding constants in the range of nM to mM.  Unlike many other methods, MST experiments can be performed rapidly and with small amounts of sample. We have two instruments for performing MST experiments, the NT.115 and the NT.LabelFree. MST experiments can be performed as a service, or you can receive training and then book the instruments yourself.

USER INFORMATION

In general, we offer four types of access to our shared research infrastructure:

  1. Research projects
  2. Full research services
  3. Trained user access | user labs
  4. Shared technology platform

Typically, these are set but not limited by the offered technology or instrument, and differ in the required user expertise, the usability of a technology, the user’s pre- and postprocessing input, and the underlying operational models.

Research project

Research projects are the equivalent of contract research organizations (CRO). The customer submits the starting material/sample and receives the ready-to-use data for publication. Hence, core facility members are often co-authoring and involved in the entire publication process.

Full research service

The user submits the sample, we perform a pre-defined workflow (incl. QC) and process the raw data. Data interpretation or contributions to publications cannot be offered in this service mode.

Trained user access | User labs

VBCF experts maintain an instrument park and train users to operate it. This requires a certain level of expertise, maturity of the offered technology, a hands-on attitude and reliability from the user.

Shared technology platform | Instrument park

These technologies require expert knowledge to run the offered instruments. Experts are hired by one of the research institutes on the Vienna BioCenter Campus. The experts maintain an instrument park, run the experiments and train other trainers. Machines can only be operated by experts.

 

The ProTech Facility provides full research service and trained user access. The ProTech Facility Usage Policy can be downloaded HERE. It is assumed that users who order services have read and understood this policy as well as the VBCF General Cooperation Conditions.

If you are a new user, please contact David Drechsel (for ProTech core services) or Arthur Sedivy (for biophysical characterization services) to discuss your project requirements. To order services and book instruments, please use our web-based booking and request system. IMP, IMBA, GMI, and MPL users can log in with their usual institute network accounts. External users must register. For questions, please contact protech(at)vbcf.ac.at.

Pricing information, additional links, information on techniques, and other documents can be found on the ProTech MyVBCF Wiki site.

We require acknowledgement of facility use in publications. 
A simple statement is sufficient and can be placed in the Materials and Methods section or in the Acknowledgments section, depending on the journal format.

Suggested format:

The XXXXXX was performed by the Protein Technologies Facility at Vienna BioCenter Core Facilities (VBCF), member of the Vienna BioCenter (VBC), Austria.

In case of (co-)authorship:

The Vienna BioCenter Core Facilities (VBCF) Protein Technologies Facility acknowledges funding from the Austrian Federal Ministry of Education, Science & Research; and the City of Vienna.

VBCF PROTEIN TECHNOLOGIES TEAM

SCIENTIFIC CONTRIBUTIONS

Amplification of human interneuron progenitors promotes brain tumors and neurological defects. Eichmüller OL, Corsini NS, Vértesy Á, Morassut I, Scholl T, Gruber VE, Peer AM, Chu J, Novatchkova M, Hainfellner JA, Paredes MF, Feucht M, Knoblich JA. Science. 2022 375(6579):eabf5546.

Panoramix SUMOylation on chromatin connects the piRNA pathway to the cellular heterochromatin machinery. Andreev VI, Yu C, Wang J, Schnabl J, Tirian L, Gehre M, Handler D, Duchek P, Novatchkova M, Baumgartner L, Meixner K, Sienski G, Patel DJ, Brennecke J. Nat Struct Mol Biol. 2022 29(2):130-142.

Sperm membrane proteins DCST1 and DCST2 are required for sperm-egg interaction in mice and fish. Noda T, Blaha A, Fujihara Y, Gert KR, Emori C, Deneke VE, Oura S, Panser K, Lu Y, Berent S, Kodani M, Cabrera-Quio LE, Pauli A, Ikawa M. Commun Biol. 2022 5(1):332.

A mitotic chromatin phase transition prevents perforation by microtubules. Schneider MWG, Gibson BA, Otsuka S, Spicer MFD, Petrovic M, Blaukopf C, Langer CCH, Batty P, Nagaraju T, Doolittle LK, Rosen MK, Gerlich DW. Nature. 2022 609(7925):183-190.

Transient upregulation of IRF1 during exit from naive pluripotency confers viral protection. Romeike M, Spach S, Huber M, Feng S, Vainorius G, Elling U, Versteeg GA, Buecker C. EMBO Rep. 2022 23(9):e55375.

A Ca2+-Mediated Switch of Epiplakin from a Diffuse to Keratin-Bound State Affects Keratin Dynamics. Ratajczyk S, Drexler C, Windoffer R, Leube RE, Fuchs P. Cells. 2022 11(19):3077.

TMBIM5 is the Ca2+ /H+ antiporter of mammalian mitochondria. Austin S, Mekis R, Mohammed SEM, Scalise M, Wang WA, Galluccio M, Pfeiffer C, Borovec T, Parapatics K, Vitko D, Dinhopl N, Demaurex N, Bennett KL, Indiveri C, Nowikovsky K. EMBO Rep. 2022 23(12):e54978

Characterization of ATG8-Family Interactors by Isothermal Titration Calorimetry. Picchianti L, Sedivy A, Dagdas Y. Methods Mol Biol. 2023;2581:149-176.

The ubiquitin ligase HOIL-1L regulates immune responses by interacting with linear ubiquitin chains. Gomez-Diaz C, Jonsson G, Schodl K, Deszcz L, Bestehorn A, Eislmayr K, Almagro J, Kavirayani A, Seida M, Fennell LM, Hagelkruys A, Kovarik P, Penninger JM, Ikeda F. iScience. 2021 24(11):103241.

PHF3 regulates neuronal gene expression through the Pol II CTD reader domain SPOC. Appel LM, Franke V, Bruno M, Grishkovskaya I, Kasiliauskaite A, Kaufmann T, Schoeberl UE, Puchinger MG, Kostrhon S, Ebenwaldner C, Sebesta M, Beltzung E, Mechtler K, Lin G, Vlasova A, Leeb M, Pavri R, Stark A, Akalin A, Stefl R, Bernecky C, Djinovic-Carugo K, Slade D. Nat Commun. 2021 12(1):6078.

Identification of lectin receptors for conserved SARS-CoV-2 glycosylation sites. Hoffmann D, Mereiter S, Jin Oh Y, Monteil V, Elder E, Zhu R, Canena D, Hain L, Laurent E, Grünwald-Gruber C, Klausberger M, Jonsson G, Kellner MJ, Novatchkova M, Ticevic M, Chabloz A, Wirnsberger G, Hagelkruys A, Altmann F, Mach L, Stadlmann J, Oostenbrink C, Mirazimi A, Hinterdorfer P, Penninger JM. EMBO J. 2021 40(19):e108375.

Molecular Characterization of the Native (Non-Linked) CD160–HVEM Protein Complex Revealed by Initial Crystallographic Analysis. Lenhartová S, Nemčovič M, Šebová R, Benko M, Zajonc DM, Nemčovičová I. Crystals. 2021; 11(7):820.

Microscale Thermophoresis and additional effects measured in NanoTemper Monolith instruments. López-Méndez B, Uebel S, Lundgren LP, Sedivy A. Eur Biophys J. 2021 50(3-4):653-660.

Applications of Golden Gate cloning to protein production using the baculovirus expression vector system. Spielvogel E, Neuhold J, Stolt-Bergner P. Methods Enzymol. 2021;660:155-169.

HUWE1 employs a giant substrate-binding ring to feed and regulate its HECT E3 domain. Grabarczyk DB, Petrova OA, Deszcz L, Kurzbauer R, Murphy P, Ahel J, Vogel A, Gogova R, Faas V, Kordic D, Schleiffer A, Meinhart A, Imre R, Lehner A, Neuhold J, Bader G, Stolt-Bergner P, Böttcher J, Wolkerstorfer B, Fischer G, Grishkovskaya I, Haselbach D, Kessler D, Clausen T.  Nat Chem Biol. 2021 doi: 10.1038/s41589-021-00831-5. Epub ahead of print.

Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study. López-Méndez B, Baron B, Brautigam CA, Jowitt TA, Knauer SH, Uebel S, Williams MA, Sedivy A, Abian O, Abreu C, Adamczyk M, Bal W, Berger S, Buell AK, Carolis C, Daviter T, Fish A, Garcia-Alai M, Guenther C, Hamacek J, Holková J, Houser J, Johnson C, Kelly S, Leech A, Mas C, Matulis D, McLaughlin SH, Montserret R, Nasreddine R, Nehmé R, Nguyen Q, Ortega-Alarcón D, Perez K, Pirc K, Piszczek G, Podobnik M, Rodrigo N, Rokov-Plavec J, Schaefer S, Sharpe T, Southall J, Staunton D, Tavares P, Vanek O, Weyand M, Wu D. Eur Biophys J. 2021 50(3-4):411-427.

Community-wide experimental evaluation of the PROSS stability-design method. Peleg Y, Vincentelli R, Collins BM, Chen KE, Livingstone EK, Weeratunga S, Leneva N, Guo Q, Remans K, Perez K, E K Bjerga G, Larsen Ø, Vaněk O, Skořepa O, Jacquemin S, Poterszman A, Kjaer S, Christodoulou E, Albeck S, Dym O, Ainbinder E, Unger T, Schuetz A, Matthes S, Bader M, de Marco A, Storici P, Semrau MS, Stolt-Bergner P, Aigner C, Suppmann S, Goldenzweig A, Fleishman SJ. J Mol Biol. 2021 Mar 26:166964.

Molecular principles of Piwi-mediated cotranscriptional silencing through the dimeric SFiNX complex. Schnabl J, Wang J, Hohmann U, Gehre M, Batki J, Andreev VI, Purkhauser K, Fasching N, Duchek P, Novatchkova M, Mechtler K, Plaschka C, Patel DJ, Brennecke J. Genes Dev. 2021 35(5-6):392-409

Temporal dissection of an enhancer cluster reveals distinct temporal and functional contributions of individual elements. Thomas HF, Kotova E, Jayaram S, Pilz A, Romeike M, Lackner A, Penz T, Bock C, Leeb M, Halbritter F, Wysocka J, Buecker C. Mol Cell. 2021 81(5):969-982

Signatures of antagonistic pleiotropy in a bacterial flagellin epitope. Parys K, Colaianni NR, Lee HS, Hohmann U, Edelbacher N, Trgovcevic A, Blahovska Z, Lee D, Mechtler A, Muhari-Portik Z, Madalinski M, Schandry N, Rodríguez-Arévalo I, Becker C, Sonnleitner E, Korte A, Bläsi U, Geldner N, Hothorn M, Jones CD, Dangl JL, Belkhadir Y. Cell Host Microbe. 2021 29(4):620-634.e9.

A complex immune response to flagellin epitope variation in commensal communities. Colaianni NR, Parys K, Lee HS, Conway JM, Kim NH, Edelbacher N, Mucyn TS, Madalinski M, Law TF, Jones CD, Belkhadir Y, Dangl JL. Cell Host Microbe. 2021 29(4):635-649

Enzyme promiscuity in natural environments: alkaline phosphatase in the ocean. Srivastava A, Saavedra DEM, Thomson B, García JAL, Zhao Z, Patrick WM, Herndl GJ, Baltar F. ISME J. 2021 doi: 10.1038/s41396-021-01013-w. Epub ahead of print.

The linear ubiquitin chain assembly complex (LUBAC) generates heterotypic ubiquitin chains. Rodriguez Carvajal A, Grishkovskaya I, Gomez Diaz C, Vogel A, Sonn-Segev A, Kushwah MS, Schodl K, Deszcz L, Orban-Nemeth Z, Sakamoto S, Mechtler K, Kukura P, Clausen T, Haselbach D, Ikeda F. Elife. 2021 10:e60660.

GoldenBac: a simple, highly efficient, and widely applicable system for construction of multi-gene expression vectors for use with the baculovirus expression vectorsystem.  Neuhold J, Radakovics K, Lehner A, Weissmann F, Queralt Garcia M, Romero MC, Berrow NS and Stolt-Bergner P [2020] BMC Biotechnology 20:26

Moyamoya disease factor RNF213 is a giant E3 ligase with a dynein-like core and a distinct ubiquitin-transfer mechanism. Ahel J, Lehner A, Vogel A, Schleiffer A, Meinhart A, Haselbach D, Clausen T [2020] eLife 9:e56185

Natural Killer Cell Activation Receptor NKp30 Oligomerization Depends on Its N-Glycosylation. Skořepa O, Pazicky S, Balousková B, Bláha J, Abreu C, Ječmen T, Rosůlek M, Fish A, Sedivy A, Harlos K, Dohnálek J, Skálová T, Vaněk O [2020] Cancers, 12(7), 1998

A PI3K-WIPI2 positive feedback loop allosterically activates LC3 lipidation in autophagy.
Fracchiolla D et a., [2020] J Cell Biol 219 (7): e201912098.
 
Cerebral organoid model reveals excessive proliferation of human caudal late interneuron progenitors in Tuberous Sclerosis Complex. Eichmüller OL et al., [2020] https://doi.org/10.1101/2020.02.27.967802  bioRxiv
 
Generation of New Isogenic Models of Huntington’s Disease Using CRISPR-Cas9 Technology.
Dabrowska M et al., [2020] J. Mol. Sci. 21(5), 1854.

A cross-kingdom conserved ER-phagy receptor maintains endoplasmic reticulum homeostasis during stress. Stephani M, Picchianti L, Gajic A, Beveridge R, Skarwan E, Sanchez de Medina Hernandez V, Mohseni A, Clavel M, Zeng Y, Naumann C, Matuszkiewicz M, Turco E, Loefke C, Li B, Dürnberger G, Schutzbier M, Chen HT, Abdrakhmanov A, Savova A, Chia KS, Djamei A, Schaffner I, Abel S, Jiang L, Mechtler K, Ikeda F, Martens S, Clausen T, Dagdas Y. Elife. 2020, 9:e58396.

Structure of the human core transcription-export complex reveals a hub for multivalent interactions. Pühringer T, Hohmann U, Fin L, Pacheco-Fiallos B, Schellhaas U, Brennecke J, Plaschka C. Elife. 2020, 9:e61503.

Site-specific ubiquitination of the E3 ligase HOIP regulates apoptosis and immune signaling. Fennell LM, Gomez Diaz C, Deszcz L, Kavirayani A, Hoffmann D, Yanagitani K, Schleiffer A, Mechtler K, Hagelkruys A, Penninger J, Ikeda F. EMBO J. 2020, 39(24):e103303.

Vaccinia Virus Immunomodulator A46: Destructive Interactions with MAL and MyD88 Shown by Negative-Stain Electron Microscopy. Azar DF, Haas M, Fedosyuk S, Rahaman MH, Hedger A, Kobe B, Skern T. Structure. 2020, 28(12):1271-1287.

Molecular mechanism of leukocidin GH-integrin CD11b/CD18 recognition and species specificity. Trstenjak N, Milić D, Graewert MA, Rouha H, Svergun D, Djinović-Carugo K, Nagy E, Badarau A. Proc Natl Acad Sci U S A. 2020 117(1):317-327.

Bridge helix arginines play a critical role in Cas9 sensitivity to mismatches. Bratovič M, Fonfara I, Chylinski K, Gálvez EJC, Sullivan TJ, Boerno S, Timmermann B, Boettcher M, Charpentier E. Nat Chem Biol. 2020 16(5):587-595.

SYNERGISTIC ON AUXIN AND CYTOKININ 1 positively regulates growth and attenuates soil pathogen resistance. Hurný A, Cuesta C, Cavallari N, Ötvös K, Duclercq J, Dokládal L, Montesinos JC, Gallemí M, Semerádová H, Rauter T, Stenzel I, Persiau G, Benade F, Bhalearo R, Sýkorová E, Gorzsás A, Sechet J, Mouille G, Heilmann I, De Jaeger G, Ludwig-Müller J, Benková E. Nat Commun. 2020, 11(1):2170.

The biophysical, molecular, and anatomical landscape of pigeon CRY4: A candidate light-based quantal magnetosensor. Hochstoeger T, Al Said T, Maestre D, Walter F, Vilceanu A, Pedron M, Cushion TD, Snider W, Nimpf S, Nordmann GC, Landler L, Edelman N, Kruppa L, Dürnberger G, Mechtler K, Schuechner S, Ogris E, Malkemper EP, Weber S, Schleicher E, Keays DA. Sci Adv. 2020, 6(33):eabb9110.

Receptor kinase module targets PIN-dependent auxin transport during canalization. Hajný J, Prát T, Rydza N, Rodriguez L, Tan S, Verstraeten I, Domjan D, Mazur E, Smakowska-Luzan E, Smet W, Mor E, Nolf J, Yang B, Grunewald W, Molnár G, Belkhadir Y, De Rybel B, Friml J. Science. 2020, 370(6516):550-557.

Human diamine oxidase cellular binding and internalization in vitro and rapid clearance in vivo are not mediated by N-glycans but by heparan sulfate proteoglycan interactions. Gludovacz E, Schuetzenberger K, Resch M, Tillmann K, Petroczi K, Vondra S, Vakal S, Schosserer M, Virgolini N, Pollheimer J, Salminen TA, Jilma B, Borth N, Boehm T. Glycobiology. 2020 cwaa090.

CRISPR-Switch regulates sgRNA activity by Cre recombination for sequential editing of two loci. Chylinski K, Hubmann M, Hanna RE, Yanchus C, Michlits G, Uijttewaal ECH, Doench J, Schramek D & Elling U. Nature Communications volume 10, Article number: 5454 (2019).

Molecular features of the UNC-45 chaperone critical for binding and folding muscle myosin.  Hellerschmied D, Lehner A, Franicevic N, Arnese R, Johnson C, Vogel A, Meinhart A, Kurzbauer R, Deszcz L, Gazda L, Geeves M & Clausen T. Nature Communications volume 10, Article number: 4781 (2019).

Data Descriptor: Map of physical interactions between extracellular domains of Arabidopsis leucine-rich repeat receptor kinases. Mott GA, Smakowska-Luzan E, Pasha A, Parys K, Howton TC, Neuhold J, Lehner A, Grünwald K, Stolt-Bergner P, Provart NJ, Mukhtar MS, Desveaux D, Guttman DS, Belkhadir Y, Scientific Data 2019, 6:190025

Transcriptional Responses to IFN-γ Require Mediator Kinase-Dependent Pause Release and Mechanistically Distinct CDK8 and CDK19 Functions. Steinparzer I, Sedlyarov V, Rubin JD, Eislmayr K, Galbraith MD, Levandowski CB, Vcelkova T, Sneezum L, Wascher F, Amman F, Kleinova R, Bender H, Andrysik Z, Espinosa JM, Superti-Furga G, Dowell RD, Taatjes DJ, Kovarik P. Molecular Cell, Volume 76, Issue 3, 7 November 2019, Pages 485-499.e8

Meiotic DNA Repair in the Nucleolus Employs a Nonhomologous End-Joining Mechanism. Sims J, Copenhaver GP, Schlögelhofer P. The Plant Cell, Vol. 31: 2259–2275, September 2019

A PI3K-WIPI2 positive feedback loop allosterically activates LC3 lipidation in autophagy. Fracchiolla D, Chang C, Hurley JH, Martens S. doi: doi.org/10.1101/2019.12.18.880591

Site-specific ubiquitination of the E3 ligase HOIP regulates cell death and immune signaling. Fennell LM, Deszcz L, Schleiffer A, Mechtler K, Kavirayani A, Ikeda F. doi: doi.org/10.1101/742544

Prdm12 Directs Nociceptive Sensory Neuron Development by Regulating the Expression of the NGF Receptor TrkA. Desiderio S, Vermeiren S, Van Campenhout C, Kricha S, Malki E, Richts S, Fletcher EV, Vanwelden T, Schmidt BZ, Henningfeld KA, Pieler T, Woods CG, Nagy V, Verfaillie C, Bellefroid EJ. Cell Rep. 2019 Mar 26;26(13): 3522-3536.

Telomerase RNAs in land plants. Fajkus P, Peška V, Závodník M, Fojtová M, Fulnečková J, Dobias Š, Kilar A, Dvořáčková M, Zachová D, Nečasová I, Sims J, Sýkorová E, Fajkus J.

The nascent RNA binding complex SFiNX licenses piRNA-guided heterochromatin formation. Batki J, Schnabl J, Wang J, Handler D, Andreev VI, Stieger CE, Novatchkova M, Lampersberger L, Kauneckaite K, Xie W, Mechtler K, Patel DJ, Brennecke J. Nat Struct Mol Biol. 2019 Aug;26(8):720-731.

A Heterochromatin-Specific RNA Export Pathway Facilitates piRNA Production. ElMaghraby MF, Andersen PR, Pühringer F, Hohmann U, Meixner K, Lendl T, Tirian L, Brennecke J. Cell. 2019 Aug 8;178(4):964-979.

Bacterial medium-chain 3-hydroxy fatty acid metabolites trigger immunity in Arabidopsis plants.
Kutschera A, Dawid C, Gisch N, Schmid C, Raasch L, Gerster T, Schäffer M, Smakowska-Luzan E, Belkhadir Y, Vlot AC, Chandler CE, Schellenberger R, Schwudke D, Ernst RK, Dorey S, Hückelhoven R, Hofmann T, Ranf S, Science. 2019 Apr 12;364(6436):178-181

Differential Action of Reelin on Oligomerization of ApoER2 and VLDL Receptor in HEK293 Cells Assessed by Time-Resolved Anisotropy and Fluorescence Lifetime Imaging Microscopy.
Dlugosz P, Tresky R, Nimpf J, Front Mol Neurosci. 2019 Feb 26;12:53

Adaptation of the Staphylococcus aureus leukocidin LukGH for the rabbit host by protein engineering.
Trstenjak N, Stulik L, Rouha H, Zmajkovic J, Zerbs M, Nagy E, Badarau A, Biochem J. 2019 Jan 25;476(2):275-292

Multiplex mutagenesis of four clustered CrRLK1L with CRISPR/Cas9 exposes their growth regulatory roles in response to metal ions
Richter J, Watson JM, Stasnik P, Borowska M, Neuhold J, Berger M, Stolt-Bergner P, Schoft V, Hauser MT. Scientific Reports, Article number: 12182 (2018)

Baculovirus-driven protein expression in insect cells: A benchmarking study
Stolt-Bergner P, Benda C, Bergbrede T, Besir H, Celie PHN, Chang C, Drechsel D, Fischer A, Geerlof A, Giabbai B, van den Heuvel J, Huber G, Knecht W, Lehner A, Lemaitre R, Nordén K, Pardee G, Racke I, Remans K, Sander A, Scholz J, Stadnik M, Storici P, Weinbruch D, Zaror I, Lua LHL, Suppmann S. J. Struct. Biol. 2018 Mar 12;

UFD-2 is an adaptor-assisted E3 ligase targeting unfolded proteins
Hellerschmied D, Roessler M, Lehner A, Gazda L, Stejskal K, Imre R, Mechtler K, Dammermann A, Clausen T. Nat Commun 2018 02 02; 9 (1)

An extracellular network of Arabidopsis leucine-rich repeat receptor kinases
Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, Layeghifard M, Neuhold J, Lehner A, Kong J, Grünwald K, Weinberger N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir Y. Nature 2018 Jan 18; 553 (7688)

qEva-CRISPR: a method for quantitative evaluation of CRISPR/Cas-mediated genome editing in target and off-target sites.
Dabrowska M, Czubak K, Juzwa W, Krzyzosiak WJ, Olejniczak M, Kozlowski P, Nucleic Acids Res. 2018 Sep 28;46(17):e101.

Cellular N-myristoyltransferases play a crucial picornavirus genus-specific role in viral assembly, virion maturation, and infectivity.
Corbic Ramljak I, Stanger J, Real-Hohn A, Dreier D, Wimmer L, Redlberger-Fritz M, Fischl W, Klingel K, Mihovilovic MD, Blaas D, Kowalski H, PLoS Pathog. 2018 Aug 6;14(8):e1007203. 

LHP1 Interacts with ATRX through Plant-Specific Domains at Specific Loci Targeted by PRC2.
Wang H, Jiang D, Axelsson E, Lorković ZJ, Montgomery S, Holec S, Pieters BJGE, Al Temimi AHK, Mecinović J, Berger F, Mol Plant. 2018 Aug 6;11(8):1038-1052.

Transcriptome Study of an Exophiala dermatitidis PKS1 Mutant on an ex Vivo Skin Model: Is Melanin Important for Infection?
Poyntner C, Mirastschijski U, Sterflinger K, Tafer H, Front Microbiol. 2018 Jul 3;9:1457.

Selective targeting of 3 repeat Tau with brain penetrating single chain antibodies for the treatment of neurodegenerative disorders.
Spencer B, Brüschweiler S, Sealey-Cardona M, Rockenstein E, Adame A, Florio J, Mante M, Trinh I, Rissman RA, Konrat R, Masliah E, Acta Neuropathol. 2018 Jul;136(1):69-87.

A Functional Study of AUXILIN-LIKE1 and 2, Two Putative Clathrin Uncoating Factors in Arabidopsis
Maciek Adamowski, Madhumitha Narasimhan, Urszula Kania, Matouš Glanc, Geert De Jaege and Jiří Friml. Plant Cell. 2018 Mar; 30(3): 700–716

Protection of Arabidopsis blunt-ended telomeres is mediated by a physical association with the Ku heterodimer
Valuchova S, Fulnecek J, Prokop Z, Stolt-Bergner P, Janouskova E, Hofr C, Riha K. Plant Cell epub 5 June 2017.

The receptor kinase FER is a RALF-regulated scaffold controlling plant immune signaling
Stegmann M, Monaghan J, Smakowska-Luzan E, Rovenich H, Lehner A, Holton N, Belkhadir Y, Zipfel C. (2017) Science Jan 20;355(6322):287-289.

Molecular basis for inner kinetochore configuration through RWD domain-peptide interactions
Schmitzberger F, Richter MM, Gordiyenko Y, Robinson CV, Dadlez M, Westermann S. EMBO J. 2017 12 01; 36 (23).

DNA Cross-Bridging Shapes a Single Nucleus from a Set of Mitotic Chromosomes
Samwer M, Schneider MWG, Hoefler R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW. Cell 2017 Aug 24; 170 (5).

HuR small molecule inhibitor elicits differential effects in adenomatosis polyposis and colorectal carcinogenesis
Lang M, Berry D, Passecker K, Mesteri I, Bhuju S, Ebner F, Sedlyarov V, Evstatiev R, Dammann K, Loy A, Kuzyk O, Kovarik P, Khare V, Beibel M, Roma G, Meisner-Kober N and Gasche C. Cancer Research epub 20 Feb 2017.

Root diffusion barrier control by a vasculature-derived peptide binding to the SGN3 receptor
Doblas VG, Smakowska-Luzan E, Fujita S, Alassimone J, Barberon M, Madalinski M, Belkhadir Y, Geldner N. (2017) Science Jan 20;355(6322):280-284.

Probing an Allosteric Pocket of CDK2 with Small Molecules
Christodoulou MS, Caporuscio F, Restelli V, Carlino L, Cannazza G, Costanzi E, Citti C, Lo Presti L, Pisani P, Battistutta R, Broggini M, Passarella D, Rastelli G. ChemMedChem 2017 Jan 05; 12 (1).

Genetic code expansion for multiprotein complex engineering
Koehler C, Sauter PF, Wawryszyn M, Girona GE, Gupta K, Landry JJ, Fritz MH, Radic K, Hoffmann JE, Chen ZA, Zou J, Tan PS, Galik B, Junttila S, Stolt-Bergner P, Pruneri G, Gyenesei A, Schultz C, Biskup MB, Besir H, Benes V, Rappsilber J, Jechlinger M, Korbel JO, Berger I, Braese S, Lemke EA. (2016). Nature Methods 13(12):997-1000 epub Oct 17, 2016

A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement
Valuchova S, Fulnecek J, Petrov AP, Tripsianes K, Riha K. (2016). Sci Rep. 6:39653.

Cross-regulation by CrcZ RNA controls anoxic biofilm formation in Pseudomonas aeruginosa
Pusic P, Tata M, Wolfinger MT, Sonnleitner E, Häussler S, Bläsi U. (2016). Sci Rep. 6:39621

Linear ubiquitination by LUBEL has a role in Drosophila heat stress response
Asaoka T, Almagro J, Ehrhardt C, Tsai I, Schleiffer A, Deszcz L, Junttila S, Ringrose L, Mechtler K, Kavirayani A, Gyenesei A, Hofmann K, Duchek P, Rittinger K, Ikeda F. (2016). EMBO Rep. 17(11):1624-1640. Epub 2016 Oct 4.

Genomic screens identify a new phytobacterial microbe-associated molecular pattern and the cognate Arabidopsis receptor-like kinase that mediates its immune elicitation
Mott AG, Thakur S, Smakowska E, Wang PW, Belkhadir Y, Desveaux D, Guttman DS. (2016). Genome Biol. 17:98.

The Sodium Glucose Cotransporter SGLT1 is an Extremely Efficient Facilitator of Passive Water Transport
Erokhova L, Horner A, Ollinger N, Siligan C, Pohl P. (2016). J Biol Chem epub March 4, 2016

The structure and regulation of human muscle alpha-actinin

Ribeiro EA Jr, Pinotsis N, Ghisleni A, Salmazo A, Konarev PV, Kostan J, Sjöblom B, Schreiner C, Polyansky AA, Gkougkoulia EA, Holt MR, Aachmann FL, Zagrović B, Bordignon E, Pirker KF, Svergun DI, Gautel M, Djinović-Carugo K. (2014). Cell 159(6):1447-60.

Structure-Function Analysis of Heterodimer Formation, Oligomerization and Receptor Binding of the Staphylococcus aureus Bi-component Toxin LukGH
Badarau A, Rouha H, Malafa S, Logan DT, Håkansson M, Stulik L, Dolezilkova I, Teubenbacher A, Gross K, Maierhofer B, Weber S, Jägerhofer M, Hoffmann D, Nagy E. (2014). J Biol Chem epub2014 Nov 3.

Assembly Mechanism of Trypanosoma brucei BILBO1, a Multidomain Cytoskeletal Protein
Vidilaseris K, Shimanovskaya E, Esson HJ, Morriswood B, Dong G (2014). J Biol Chem 289(34):23870-81.

A strand-specific switch in noncoding transcription switches the function of a Polycomb/Trithorax response element
Herzog VA, Lempradl A, Trupke J, Okulski H, Altmutter C, Ruge F, Boidol B, Kubicek S, Schmauss G, Aumayr A, Ruf M, Pospisilik A, Dimond A, Senergin HB, Vargas ML, Simon JA, Ringrose L. (2014). Nat Genet 46(9):973-981.

The unique regulation of iron-sulfur cluster biogenesis in a Gram-positive bacterium
Santos JA, Alonso-García N, Macedo-Ribeiro S, Pereira PJ. (2014). Proc Natl Acad Sci 111(22):E2251-60.

Characterization and Structure of the Vaccinia Virus NF-kB Antangonist A46
Fedosyuk, S., Grishkovskaya I., de Almeida Ribeiro E Jr., Skern T. (2014). J Biol Chem 289(6):3749-62.

Flexible long-range loops in the VH gene region of the Igh locus facilitate the generation of a diverse antibody repertoire
Medvedovic J, Ebert A, Tagoh H, Tamir IM, Schwickert TA, Novatchkova M, Sun Q, Huis In't Veld PJ, Guo C, Yoon HS, Denizot Y, Holwerda SJ, de Laat W, Cogne M, Shi Y, Alt FW, Busslinger M (2013). Immunity 39(2):229-44.

Sufficient amounts of functional HOP2/MND1 complex promote interhomolog DNA repair but are dispensable for intersister DNA repair during meiosis in Arabidopsis
Uanschou C, Roncerat A, Von Harder M, De Muyt A, Vezon D, Pereira L, Chelysheva L, Kobayashi W, Kurumizaka H, Schloegelhofer P, Grelon M (2013).
Plant Cell 25(12):4924-40.