Renata Grzywa1, Ewa Gorodkiewicz2, Ewa Burchacka1, Adam Lesner3, Piotr Laudański4, Zenon Łukaszewski5, Marcin Sieńczyk6. 1. Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland. 2. Department of Electrochemistry, Institute of Chemistry, University of Bialystok, Al. J. Piłsudskiego 11/4, 15-443 Bialystok, Poland. 3. Faculty of Chemistry, University of Gdansk, ul. Sobieskiego 18, 80-952 Gdansk, Poland. 4. Department of Perinatology and Obstetrics, Medical University of Bialystok, ul. Marii Sklodowskiej-Curie 24a, 15-276 Bialystok, Poland. 5. Institute of Chemistry, Poznan University of Technology, Piotrowo 3, 60-965 Poznan, Poland. 6. Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland. Electronic address: marcin.sienczyk@pwr.wroc.pl.
Abstract
OBJECTIVE: Cathepsin G is a serine peptidase whose physiological role is mainly associated with an early immune response, anti-microbial activity as well as platelet activation or hydrolysis of coagulation factors. In addition, since the activity of cathepsin G has been associated with the development of various pathological disorders, the measurement of its activity in patient samples is of high interest. Unfortunately, the usefulness of common immunological methods is limited, since they cannot distinguish between catalytically active and inactive protease. STUDY DESIGN: Here we present the application of recently developed Surface Plasmon Resonance-based biosensor for the detection of active cathepsin G in human endometrium samples. The key element of the system is based on the irreversible binding of cathepsin G to its specific phosphonic-type inhibitor immobilized on the surface of the gold chip. The concentration of cathepsin G was measured in tissue samples from the group of patients with endometriosis as well as in the control group. RESULTS: The level of cathepsin G ascertained in endometrium tissue samples was over twice as high for the group of patients suffering from endometriosis as compared to the control group, with the median values of 0.5 pmol/mg and 0.2 pmol/mg, respectively. CONCLUSION: The SPR sensor armed with a specific irreversible phosphonic inhibitor represents a highly useful tool for the determination of catalytically active cathepsin G concentration in endometrial tissue.
OBJECTIVE:Cathepsin G is a serine peptidase whose physiological role is mainly associated with an early immune response, anti-microbial activity as well as platelet activation or hydrolysis of coagulation factors. In addition, since the activity of cathepsin G has been associated with the development of various pathological disorders, the measurement of its activity in patient samples is of high interest. Unfortunately, the usefulness of common immunological methods is limited, since they cannot distinguish between catalytically active and inactive protease. STUDY DESIGN: Here we present the application of recently developed Surface Plasmon Resonance-based biosensor for the detection of active cathepsin G in human endometrium samples. The key element of the system is based on the irreversible binding of cathepsin G to its specific phosphonic-type inhibitor immobilized on the surface of the gold chip. The concentration of cathepsin G was measured in tissue samples from the group of patients with endometriosis as well as in the control group. RESULTS: The level of cathepsin G ascertained in endometrium tissue samples was over twice as high for the group of patients suffering from endometriosis as compared to the control group, with the median values of 0.5 pmol/mg and 0.2 pmol/mg, respectively. CONCLUSION: The SPR sensor armed with a specific irreversible phosphonic inhibitor represents a highly useful tool for the determination of catalytically active cathepsin G concentration in endometrial tissue.
Authors: A Sankiewicz; L Romanowicz; P Laudanski; B Zelazowska-Rutkowska; B Puzan; B Cylwik; E Gorodkiewicz Journal: Anal Bioanal Chem Date: 2016-05-21 Impact factor: 4.142