Winners of Foundation for Polish Science programmes from the Wrocław University of Technology, in cooperation with American researchers, have developed a new method for parallel imaging of the activity of enzymes. The results of the research have just been published in the prestigious Journal of the American Chemical Society. The first author of the work is Dr Paulina Kasperkiewicz, a two-time winner of FNP’s START programme, and the other Polish co-author is Prof. Marcin Drąg, also a winner of the START programme as well as the FOCUS programme.
Proteases are enzymes controlling practically all metabolic pathways in living organisms. Disruption of their activity leads to development of many civilizational diseases, including cancer, diabetes, neurodegenerative illnesses and loss of resistance to pathogens. In recent years, many important questions about the involvement of proteases in the development of specific diseases have been successfully answered using chemical markers (activity-based probes). An advantage of using such markers is their ability to depict only active enzymes, that is, those that take an active part in a given biological process. Nonetheless, one of the great remaining challenges is to create markers acting selectively with one enzyme, and the possibility of imaging several enzymes in one place at the same time.
In the article in JACS, “A toolbox of fluorescent probes for parallel imaging reveals uneven location of serine proteases in neutrophils ,” the researchers from the Technical University of Wrocław, in cooperation with researchers from the Sanford Burnham Prebys Medical Discovery Institute in La Jolla, California, for the first time largely succeeded in solving these problems. They designed and synthesized chemical markers with very high specificity and additionally equipped with various fluorophores. The arrangement they called “a toolbox of fluorescent probes” was used to image the activity of serine proteases in neutrophils—enzymes particularly important in protecting the host organism against pathogens but also involved in the development of cancers, primarily of the lung. Using methods of imaging with a fluorescent microscope and flow cytometry, they succeeded in showing that parallel monitoring of neutrophil enzymes at the same time is possible despite the different activity and concentration in the neutrophils. Moreover, using these probes, the researchers demonstrated uneven distribution of neutrophil serine proteases in neutrophil azurophil granules. This mutual exclusion between serine proteases in neutrophils was never observed before, suggesting the existence of unknown mechanisms for targeting these enzymes to granules. This observation may have great biological significance, and the research is continuing.
The lead author for the entire project is Dr Paulina Kasperkiewicz, who led the research in the team of Prof. Marcin Drąg and in a postdoctoral fellowship in the team of Prof. Guy S. Salvesen (Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California).
Our hearty congratulations!
Pictured: Dr Paulina Kasperkiewicz and Prof. Marcin Drąg. Photo: private archive