title: Design and synthesis of new substrates of HtrA2 protease.
abstract: HtrA2 belongs to the HtrA (high temperature requirement A) family of ATP-independent serine proteases. The primary function of HtrA2 includes maintaining the mitochondria homeostasis, cell death (by apoptosis, necrosis, or anoikis), and contribution to the cell signaling. Several recent reports have shown involvement of HtrA2 in development of cancer and neurodegenerative disorders. Here, we describe the profiling of HtrA2 protease substrate specificity via the combinatorial chemistry approach that led to the selection of novel intramolecularly quenched substrates. For all synthesized compounds, the highest HtrA2-mediated hydrolysis efficiency and selectivity among tested HtrA family members was observed for ABZ-Ile-Met-Thr-Abu-Tyr-Met-Phe-Tyr(3-NO2)-NH2, which displayed a specificity constant kcat/KM value of 14,535M(-1)s(-1).
authors: Faculty of Chemistry, University of Gdansk, 80-308 Gdansk, Poland. Electronic address: magdalena.wysocka@ug.edu.pl.
authors: Magdalena
authors: M
authors: Wysocka
authors: Faculty of Chemistry, University of Gdansk, 80-308 Gdansk, Poland.
authors: Anna
authors: A
authors: Wojtysiak
authors: Faculty of Chemistry, University of Gdansk, 80-308 Gdansk, Poland.
authors: Małgorzata
authors: M
authors: Okońska
authors: Faculty of Chemistry, University of Gdansk, 80-308 Gdansk, Poland.
authors: Natalia
authors: N
authors: Gruba
authors: Faculty of Biology, University of Gdansk, 80-308 Gdansk, Poland.
authors: Mirosław
authors: M
authors: Jarząb
authors: Faculty of Biology, University of Gdansk, 80-308 Gdansk, Poland.
authors: Tomasz
authors: T
authors: Wenta
authors: Faculty of Biology, University of Gdansk, 80-308 Gdansk, Poland.
authors: Barbara
authors: B
authors: Lipińska
authors: Faculty of Chemistry, Wroclaw Technical University, 50-370 Wrocław, Poland.
authors: Reneta
authors: R
authors: Grzywa
authors: Faculty of Chemistry, Wroclaw Technical University, 50-370 Wrocław, Poland.
authors: Marcin
authors: M
authors: Sieńczyk
authors: Faculty of Chemistry, University of Gdansk, 80-308 Gdansk, Poland.
authors: Krzysztof
authors: K
authors: Rolka
authors: Faculty of Chemistry, University of Gdansk, 80-308 Gdansk, Poland.
authors: Adam
authors: A
authors: Lesner
mesh_terms: D000074923:High-Temperature Requirement A Serine Peptidase 2; D006801:Humans; D024101:Mitochondrial Proteins / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D012697:Serine Endopeptidases / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D024101:Mitochondrial Proteins; D010455:Peptides; D012697:Serine Endopeptidases; C000615914:HTRA2 protein, human; D000074923:High-Temperature Requirement A Serine Peptidase 2
keywords: Fluorogenic substrates; HtrA2 proteinase; Peptide libraries; Proteolytic enzymes