title: Virtual screening of novel reversible inhibitors for marine alkaline protease MP.
abstract: Marine alkaline protease (MP,(2) accession no. ACY25898) is produced by a marine bacterium strain isolated from Yellow Sea sediment in China. Previous research has shown that this protease is a cold-adapted enzyme with antioxidant activity that could be used as a detergent additive. Owing to its instability in the liquid state, MP's application in liquid detergents was limited. Therefore, the discovery of reversible MP inhibitors to stabilize the protease was imperative. Here, we used the X-ray structure of MP and recompiled AutoDock 4.2 with refined Zn(2+) characters to screen the free chemical database ZINC. After completing the docking procedure, we applied strategies including the "initial filter", consensus scoring and pharmocophore model to accelerate the process and improve the virtual screening success rate. The "initial filter" was built based on the docking results of boronic acid derivatives validated as reversible inhibitors of MP by our previous studies. Finally, ten compounds were purchased or synthetized to test their binding affinity for MP. Three of the compounds could reversibly inhibit MP with apparent Ki values of 0.8-1.2 mmol. These active compounds and their binding modes provide useful information for understanding the molecular mechanism of reversible MP inhibition. The results may also serve as the foundation for further screening and design of reversible MP inhibitors.
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Qingdao Key Laboratory of Marine Enzyme Engineering, Qingdao 266071, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Ji
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zheng
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Sheng
authors.forename: Jianhua
authors.initials: J
authors.lastname: Hao
authors.forename: Mi
authors.initials: M
authors.lastname: Sun
mesh_terms: D020816:Amino Acid Motifs; D059001:Aquatic Organisms; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D020134:Catalytic Domain; D004353:Drug Evaluation, Preclinical; D010450:Endopeptidases / Q000737:chemistry*; D004795:Enzyme Stability; D005417:Flavobacterium / Q000201:enzymology; D062105:Molecular Docking Simulation; D011480:Protease Inhibitors / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D011480:Protease Inhibitors; D010450:Endopeptidases; C087822:alkaline protease
keywords: Marine alkaline protease; Reversible inhibitor; Virtual screening