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Published on: October 1, 2016
Optimization of fluorescent substrates for ADAM17 and their utility in the detection of diabetes
Honorata Sikora1, Natalia Gruba2, Magdalena Wysocka1
1Department of Biomedical Chemistry, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63, 80-308, Gdańsk, Poland.
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.
Insights
Researchers developed a new method to detect ADAM17 activity in diabetic patients. This enzyme
Area of Science:
- Biochemistry
- Enzymology
- Nephrology
Background:
- ADAM17 (a disintegrin and metalloproteinase 17) is a sheddase enzyme.
- ADAM17 is implicated in the cleavage of the antiaging protein Klotho (KL).
- Reduced Klotho levels are linked to early chronic kidney disease stages.
Purpose of the Study:
- To optimize specific and selective fluorescent substrates for ADAM17.
- To utilize the optimized substrate for detecting ADAM17 activity in patient urine samples.
Main Methods:
- Development and optimization of ADAM17-specific fluorescent substrates.
- Assay development for detecting proteolytic activity in biological samples.
- Analysis of urine samples from diabetic patients and healthy individuals.
Main Results:
- Successfully optimized ADAM17-specific fluorescent substrates.
- Detected significant ADAM17 proteolytic activity in urine samples from all tested diabetic patients.
- No detectable ADAM17 activity was observed in urine samples from healthy individuals.
Conclusions:
- The developed assay can detect ADAM17 activity in urine.
- Elevated ADAM17 activity may be a biomarker for diabetes-related kidney complications.
- Further research is warranted to explore the clinical utility of this finding.
