Development of chemical tools to monitor human kallikrein 13 (KLK13) activity

Natalia Gruba , Ewa Bielecka , Magdalena Wysocka , Anna Wojtysiak , Magdalena Brzezińska-Bodal , Kamila Sychowska , Magdalena Kalińska , Małgorzata Magoch , Aleksandra Pęcak , Katherine Falkowski , Magdalena Wiśniewska , Laura Sąsiadek , Karolina Płaza , Eileen Kroll , Anastasija Pejkovska , Maren Rehders , Klaudia Brix , Grzegorz Dubin , Tomasz Kantyka , Jan Potempa , Adam Lesner


Kallikrein 13 (KLK13) was first identified as an enzyme that is downregulated in a subset of breast tumors. This serine protease has since been implicated in a number of pathological processes including ovarian, lung and gastric cancers. Here we report the design, synthesis and deconvolution of libraries of internally quenched fluorogenic peptide substrates to determine the specificity of substrate binding subsites of KLK13 in prime and non-prime regions (according to the Schechter and Berger convention). The substrate with the consensus sequential motive ABZ-Val-Arg-Phe-Arg-ANB-NH2 demonstrated selectivity towards KLK13 and was successfully converted into an activity-based probe by the incorporation of a chloromethylketone warhead and biotin bait. The compounds described may serve as suitable tools to detect KLK13 activity in diverse biological samples, as exemplified by overexpression experiments and targeted labeling of KLK13 in cell lysates and saliva. In addition, we describe the development of selective activity-based probes targeting KLK13, to our knowledge the first tool to analyze the presence of the active enzyme in biological samples.
Author Natalia Gruba (FCh / DET / LBAN)
Natalia Gruba,,
- Laboratory of Biochemical Analytics and Nanodiagnostics
, Ewa Bielecka
Ewa Bielecka,,
, Magdalena Wysocka (FCh / DBCh / LMCh)
Magdalena Wysocka,,
- Laboratory of Medical Chemistry
, Anna Wojtysiak (FCh / DMB / LBA)
Anna Wojtysiak,,
- Laboratory of Biochemical Analytics
, Magdalena Brzezińska-Bodal (FCh / DMB / LBA)
Magdalena Brzezińska-Bodal,,
- Laboratory of Biochemical Analytics
, Kamila Sychowska (FCh / DMB / LBA)
Kamila Sychowska,,
- Laboratory of Biochemical Analytics
, Magdalena Kalińska
Magdalena Kalińska,,
, Małgorzata Magoch
Małgorzata Magoch,,
, Aleksandra Pęcak
Aleksandra Pęcak,,
, Katherine Falkowski
Katherine Falkowski,,
et al.`
Journal seriesInternational Journal of Molecular Sciences, ISSN 1422-0067, (A 30 pkt)
Issue year2019
Keywords in EnglishKLK13, substrate specificity, combinatorial chemistry, fluorogenic substrate, activity-based probe
ASJC Classification1604 Inorganic Chemistry; 1605 Organic Chemistry; 1606 Physical and Theoretical Chemistry; 1706 Computer Science Applications; 1607 Spectroscopy; 1312 Molecular Biology; 2700 General Medicine; 1503 Catalysis
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)30
ScoreMinisterial score = 30.0, 24-07-2019, ArticleFromJournal
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2016 = 1.147; WoS Impact Factor: 2017 = 3.687 (2) - 2017=3.878 (5)
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