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
AbstractKallikrein 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.
|Journal series||International Journal of Molecular Sciences, ISSN 1422-0067, (N/A 100 pkt)|
|Publication size in sheets||0.85|
|Keywords in English||KLK13, substrate specificity, combinatorial chemistry, fluorogenic substrate, activity-based probe|
|ASJC Classification||; ; ; ; ; ; ;|
|License||Journal (articles only); published final; ; with publication|
|Score||= 100.0, 10-01-2020, ArticleFromJournal|
|Publication indicators||= 1; = 2; : 2018 = 1.224; : 2018 = 4.183 (2) - 2018=4.331 (5)|
|Citation count*||2 (2020-01-15)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.