Laboratory of Comparative Enzymology

Head: Wojciech RODE

Staff: Joanna Cieśla, Magdalena Dąbrowska, Anna Dowierciał, Adam Jarmuła, Piotr Maj (PhD student), Piotr Wilk (PhD student), Zbigniew Zieliński

Research profile:

Different aspects of thymidylate biosynthesis, such as its enzymology, structure, regulation, inhibition, as well as post- translational modifications, and their influence on the enzyme properties are investigated. Problems connected with thymidylate synthase being a target in chemotherapy, including search for new inhibitors/drugs, drug resistance, specificity and mechanism of action, as well as mechanisms of enzyme inhibition and its specificity, are of particular interest. Another line of studies focuses on cell fate decision mechanisms, using two model experimental systems of senescence-like growth arrest, the nurse cell of Trichinella spiralis parasite and metho- trexate-exposed human colon cancer cells, with bioinformatic approach applied to infer active signaling pathways. Certain molecular aspects of host immunological response to Trichinella spiralis infection are also studied

Current research activities:

• developmental pattern of expression of enzymes involved in nematode thymidylate biosynthesis as potential new target for antiparasitic chemotherapy
• effects of post-translational modifications on thymidylate synthase catalytic and non-catalytic (binding repressing translation of its own mRNA) properties.
• mechanism of thymidylate synthase reaction and its inhi- bition, including search for new inhibitors
• mechanism of establishment of methotrexate–induced cancer cell senescence

Methods:

• classic enzymology, incl. kinetic studies
• genomics and genetic engineering
• purified recombinant enzyme protein production
• enzyme protein chemical modification
• molecular modeling (molecular dynamics, quantum mechanics)
• protein crystallography

Selected publications:

Dąbrowska-Maś E., Frączyk T., Ruman T., Radziszewska K., Wilk P., Cieśla J., Zieliński Z., Jurkiewicz A., Gołos B., Wińska P., Wałajtys-Rode E., Leś A., Nizioł J., Jarmuła A., Stefanowicz P., Szewczuk Z., Rode W. (2012) Tyrosine nitration affects thymi- dylate synthase properties. Organic & Biomolecular Chemistry, 10: 323-331.

Gołos B., Dąbrowska M., Wałajtys-Rode E., Zieliński Z., Wińska P., Cieśla J., Jagielska E., Moczoń T., Rode W. (2012) Immunofluorescent localization of thymidylate synthase in the development of Trichinella spiralis and Caenorhabditis elegans. Molecular and Biochemical Parasitology, 183: 63-69.

Dąbrowska M., Skoneczny M., Rode W. (2011) Functional gene expression profile underlying methotrexate-induced senes- cence in human colon cancer cells. Tumor Biology, 32: 965–976.

Jarmuła A., Frączyk T., Cieplak P., Rode W. (2010) Mechanism of influence of phosphorylation on serine 124 on a decrease of catalytic activity of human thymidylate synthase. Bioorganic & Medicinal Chemistry, 18: 3361-3370.

Świniarska M., Leś A., Rode W., Cieśla J., Millán-Pacheco C., Blake I.O., Pastor N. (2010) Segmental motions of rat thymidylate synthase leading to half-the-sites Behaviour. Biopolymers, 93: 549-559