Protein Engineering Laboratory

Dr. Anna Urbanowicz

Head of Laboratory
aniau@ibch.poznan.pl
ext. 1152

Paulina Bierwagen MSc 
Phd student

PhD students

Dr. Jakub Barciszewski
specialist biologist

Alina Kasperska MSc
specjalist biologist

Technical Staff:

Dr. habil. Michał Sikorski

associate professor

Dr. Joanna Śliwiak

adiunkt

Dr. Marta Grzechowiak

adiunkt

Researchers:

  • J. Smietanska, J. Sliwiak, M. Gilski, Z. Dauter, R. Strzalka, J. Wolny, M. Jaskolski, A new modulated crystal structure of the ANS complex of the St John's wort Hyp-1 protein with 36 protein molecules in the asymmetric unit of the supercell. Acta cryst. D. 2020, 76, 653-667 

  • M. Grzechowiak, J. Sliwiak, M. Jaskolski, M. Ruszkowski, Structural studies of glutamate dehydrogenase (Isoform 1) from Arabidopsis thaliana, an important enzyme at the branch-point between carbon and nitrogen metabolism. Front. Plant Sci. 2020, 11,  754 

  • M. Grzechowiak, M. Ruszkowski, J. Sliwiak, K. Szpotkowski, M. Sikorski, M. Jaskolski, Crystal structures of plant inorganic pyrophosphatase, an enzyme with a moonlighting autoproteolytic activity Biochemical Journal 2019, 476, 2297-2319

  • P. Bierwagen, K. Szpotkowski, M. Jaskolski, A. Urbanowicz, Borrelia outer surface protein C is capable of human fibrinogen binding FEBS Journal 2019, 286, 2415-2428

  • J. Czyrko, B. Imiolczyk, J. Sliwiak, K. Szpotkowski, J.F. Barciszewski, M. Jaskolski, K. Brzezinski, Biochemical and structural characterization of an unusual cyanobacterial S-adenosyl-L-homocysteine hydrolase from Synechocystis sp. PCC 6803, 44th FEBS Congress 2019. FEBS OPEN BIO 2019, 9 (S.1), 267-267

  • B. Shashikadze, M. Grzechowiak, M. Jaskolski, B. Marciniak, M. Ignasiak Kciuk, Iodide prevents proteins from oxidative damage. From traditional biochemical methods to crystallographic tools, Annual Meeting of the Society-for-Free-Radical-Research- Europe (SFRRE) on Redox 2019 FREE RADICAL BIOLOGY AND MEDICINE  2019, 139 (S.1), 167

  • Belter, K. Szpotkowski, M. Popenda, J. F. Barciszewski, Pioneering structural and functional studies of miRNA, 44th FEBS Congress 2019 FEBS OPEN BIO 2019, 9 (S.1), 243-244

  • J. Sliwiak, M. Sikorski, M. Jaskolski, PR-10 proteins as potential mediators of melatonin-cytokinin cross-talk in plants: crystallographic studies of LlPR-10.2B isoform from yellow lupine FEBS Journal 2018, 285, 1907-1922

  • J. Czyrko, J. Sliwiak, B. Imiolczyk, Z. Gdaniec, M. Jaskolski, K. Brzezinski, Metal-cation regulation of enzyme dynamics is a key factor influencing the activity of S-adenosyl-L-homo¬cysteine hydrolase from Pseudomonas aeruginosa Scientific Reports 2018, 8, 11334

  • A.A. Mieloch, M. Krecisz, J.D. Rybka, A. Strugala, M. Krupinski, A. Urbanowicz, M. Kozak, B. Skalski, M. Figlerowicz, M. Giersig, The influence of ligand charge and length on the assembly of Brome mosaic virus derived virus-like particles with magnetic core AIP Advances 2018, 8, 035005

  • A. Strugala, P. Bierwagen, J.D. Rybacka, M. Giersig, M. Figlerowicz, A. Urbanowicz; BMV propagation, extraction and purification using chromatographic methods Bio-Protocol 2018, 8

  • K. Brzezinski, J. Czyrko, J. Sliwiak, E. Nalewajko-Sieliwoniuk, M. Jaskolski, B. Nocek, Z. Dauter, S-adenosyl-L-homocysteine hydrolase from a hyperthermophile (Thermatoga maritima) is expressed in Escherichia coli in inactive form - biochemical and structural studies International Journal of Biological Macromolecules 2017, 104, 584-596 

  • A. Strugala, M. Krecisz, J.D. Rybka, A. Urbanowicz, K. Szpotkowski, P. Bierwagen, M. Figlerowicz, M. Kozak, C. Böttcher, M. Giersig, Biophysical analysis of BMV virions purified using a novel method Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences 2017, 1068-1069, 157-163

  • M. Jaskolski, J. Sliwiak, The phytohormone ligand repertoire of plant PR-10 proteins includes melatonin FEBS Journal 2017, 284, 34

Selected publications

  • Structural and functional studies of proteins crucial for the interactions between ticks, vertebrates and Borrelia.  

  • Obtaining PR-10 protein isoforms from St. John's woofer as potential melatonin receptors and mediators for hormonal signal transmission, and preliminary crystallographic characteristics of their complexes with natural ligands. NCN- MINIATURE 2 

  • Characteristics of virus-like particles formed on the basis of the recombined capsid protein from plant icosahedral virus.

Research activity

  • a set of expression vectors and equipment necessary for cloning and production of proteins in the bacterial system,

  • incubator with shaking and temperature control in the range of 5 to 50ºC,

  • sonicator with variable probes depending on sample volume,

  • preparative centrifuge (rotors: 8 x 50 ml and 6 x 500 ml),

  • vacuum set for the affinity chromatography,

  • fast protein liquid chromatography system (FPLC) equipped with size exclusion columns (SEC),

  • analyzer Zetasizer µV Malvern for static and dynamic light scattering measurements (DLS and SLS) in order to assess molecular mass and hydrodynamic diameter of proteins and polymers,

  • calorimeters Microcal iTC200 Malvern and Microcal PEAQ-ITC Malvern for assessing the parameters of ligand binding using isothermal titration calorimetry (ITC),

  • system Monolith NT.115 Nanotemper for assessing the parameters of ligand binding using microscale thermophoresis (MST),

  • system Octet K2 ForteBio for assessing the parameters of ligand binding using biolayer interferometry (BLI), 

  • nanophotometer N60 Implen, 

  • spectrometer Agilent 8453 UV-visible for concentration and kinetics measurements,

  • crystallization robot ARI Gryphon and commercial sets of reagents from Hampton Research, MDL, for initial crystallization conditions screening and optimization of the conditions via vapor diffusion method,

  • diffractometer Rigaku XtaLAB Synergy-R for crystals quality testing and diffraction patterns recording,

  • cold-room (8 ºC) and crystallization room (18 ºC).

Equipment

We offer our equipment and provide expertise in terms of preparation of expression vectors, production of recombinant proteins and their purification, as well as evaluation of quality of protein preparations intended for functional and structural research. 

We have experience and equipment enabling physicochemical characteristics of interactions between proteins and other macromolecules. Our measurement methods allow not only to assess affinity and measure constants of association and dissociation, but also to determine the number of ligand binding sites or thermodynamic parameters of binding such as enthalpy or entropy. 

We conduct high-throughput tests of the initial conditions for crystallization of biomolecules as well as optimization of the crystallization process to obtain crystals for diffraction experiments. We record diffraction data for protein crystals and nucleic acids using a diffractometer equipped with a rotating anode.

Offer

  • cloning, mutagenesis, expression plasmids

  • protein production in bacterial system

  • isothermal titration calorimetry (ITC)

  • dynamic and static light scattering (DLS, SLS)

  • microscale thermophoresis (MST)

  • biolayer interferometry (BLI)

  • fast protein liquid chromatography (FPLC)

  • protein and nucleic acids crystallization

  • X-ray diffraction measurements

  • macromolecules Structure solving

  • Ixodes ticks and Borrelia spirochetes proteins

  • complexes of pathogenesis related class 10 plant protein (PR-10)

  • fitomelatonin 

  • transcription factors WRKY

  • plant pyrophosphatase AtPPA1

  • viral capsids assembly

  • plant glutamate dehydrogenase

Keywords

Research Projects

Media Społecznościowe

Strona zrobiona w kreatorze stron internetowych WebWave

Cooperation

Institute of Bioorganic Chemistry,
Polish Academy of Sciences
Z. Noskowskiego 12/14
61-704 Poznań
tel centrala: (+48) 61 852 85 03
fax: (+48) 61 852 05 32
e-mail: ibch@ibch.poznan.pl

Sekretariat Dyrektora, tel: 61 852 89 19

Adress