Optimization of fluorescent substrates for ADAM17 and their utility in the detection of diabetes

Honorata Sikora; Natalia Gruba; Magdalena Wysocka; Agnieszka Piwkowska; Adam Lesner

doi:10.1016/j.ab.2023.115337

Optimization of fluorescent substrates for ADAM17 and their utility in the detection of diabetes

Anal Biochem. 2023 Nov 15:681:115337. doi: 10.1016/j.ab.2023.115337. Epub 2023 Sep 30.

Authors

Honorata Sikora¹, Natalia Gruba², Magdalena Wysocka¹, Agnieszka Piwkowska³, Adam Lesner⁴

Affiliations

¹ Department of Biomedical Chemistry, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63, 80-308, Gdańsk, Poland.
² Department of Environmental Technology, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63 Street, PL, 80-308, Gdańsk, Poland. Electronic address: natalia.gruba@ug.edu.pl.
³ Laboratory of Molecular and Cellular Nephrology, Mossakowski Medical Research Institute Polish Academy of Sciences, Wita Stwosza 63, 80-308, Gdansk, Poland; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63, 80-308, Gdansk, Poland.
⁴ Department of Environmental Technology, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63 Street, PL, 80-308, Gdańsk, Poland.

PMID: 37783443
DOI: 10.1016/j.ab.2023.115337

Abstract

ADAM17 (a disintegrin and metalloproteinase 17) is a sheddase that releases various types of membrane-associated proteins, including adhesive molecules, cytokines and their receptors, and inflammatory mediators. Evidence suggests that the enzyme is involved in the proteolytic cleavage of antiaging transmembrane protein Klotho (KL). What is more, reduced serum and urinary KL levels are observed in the early stages of chronic kidney disease. This study aimed to optimise the ADAM17 specific and selective fluorescent substrates. Then, the obtained substrate was used to detect the enzyme in urine samples of patients diagnosed with diabetes. It turned out that in all cases we were able to detect proteolytic activity, which was the opposite of the healthy samples.

Keywords: ADAM17; Combinatorial chemistry; Diabetes; Fluorogenic peptides.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ADAM17 Protein
Diabetes Mellitus* / diagnosis
Humans
Membrane Proteins
Proteolysis

Substances

ADAM17 Protein
Membrane Proteins
ADAM17 protein, human