title: Chemoproteomic, biochemical and pharmacological approaches in the discovery of inhibitors targeting human α/β-hydrolase domain containing 11 (ABHD11).
abstract: ABHD11 (α/β-hydrolase domain containing 11) is a non-annotated enzyme belonging to the family of metabolic serine hydrolases (mSHs). Its natural substrates and products are unknown. Using competitive activity-based protein profiling (ABPP) to identify novel inhibitors of human (h)ABHD11, three compounds from our chemical library exhibited low nanomolar potency towards hABHD11. Competitive ABPP of various proteomes revealed fatty acid amide hydrolase (FAAH) as the sole off-target among the mSHs. Our fluorescent activity assays designed for natural lipase substrates revealed no activity of hABHD11 towards mono- or diacylglycerols. A broader profiling using para-nitrophenyl (pNP)-linked substrates indicated no amidase/protease, phosphatase, sulfatase, phospholipase C or phosphodiesterase activity. Instead, hABHD11 readily utilized para-nitrophenyl butyrate (pNPC4), indicating lipase/esterase-type activity that could be exploited in inhibitor discovery. Additionally, a homology model was created based on the crystal structure of bacterial esterase YbfF. In contrast to YbfF, which reportedly hydrolyze long-chain acyl-CoA, hABHD11 did not utilize oleoyl-CoA or arachidonoyl-CoA. In conclusion, the present study reports the discovery of potent hABHD11 inhibitors with good selectivity among mSHs. We developed substrate-based activity assays for hABHD11 that could be further exploited in inhibitor discovery and created the first homology-based hABHD11 model, offering initial insights into the active site of this poorly characterized enzyme.
authors.affiliation: School of Medicine, Institute of Biomedicine, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland. Electronic address: dina.navia-paldanius@uef.fi.
authors.forename: Dina
authors.initials: D
authors.lastname: Navia-Paldanius
authors.affiliation: School of Pharmacy, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Jayendra Z
authors.initials: JZ
authors.lastname: Patel
authors.affiliation: School of Medicine, Institute of Biomedicine, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Miriam
authors.initials: M
authors.lastname: López Navarro
authors.affiliation: School of Medicine, Institute of Biomedicine, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Hermina
authors.initials: H
authors.lastname: Jakupović
authors.affiliation: Department of Biology, University of Eastern Finland, P.O Box 111, 80101 Joensuu, Finland.
authors.forename: Steffi
authors.initials: S
authors.lastname: Goffart
authors.affiliation: School of Medicine, Institute of Biomedicine, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Sanna
authors.initials: S
authors.lastname: Pasonen-Seppänen
authors.affiliation: School of Pharmacy, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Tapio J
authors.initials: TJ
authors.lastname: Nevalainen
authors.affiliation: School of Medicine, Institute of Biomedicine, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland; Institute of Dentistry, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Tiina
authors.initials: T
authors.lastname: Jääskeläinen
authors.affiliation: School of Pharmacy, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Tuomo
authors.initials: T
authors.lastname: Laitinen
authors.affiliation: School of Medicine, Institute of Biomedicine, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Jarmo T
authors.initials: JT
authors.lastname: Laitinen
authors.affiliation: School of Medicine, Institute of Biomedicine, University of Eastern Finland, P.O Box 1627, 70211 Kuopio, Finland.
authors.forename: Juha R
authors.initials: JR
authors.lastname: Savinainen
mesh_terms: D000818:Animals; D005558:Arylformamidase / Q000235:genetics; D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D055808:Drug Discovery; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000378:metabolism; D008958:Models, Molecular; D040901:Proteomics; D057057:Serine Proteases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013869:Thiolester Hydrolases / Q000235:genetics
chemical_list: D015842:Serine Proteinase Inhibitors; C581317:LYPLA2 protein, human; D013869:Thiolester Hydrolases; C568817:ABHD11 protein, human; D057057:Serine Proteases; D005558:Arylformamidase
keywords: ABHD11; ABPP; AFMID; Homology modelling; Inhibitor screening; LYPLA2; Serine hydrolases