Rat High-temperature requirement factor A3 ELISA Kit

Human HtrA3 (High temperature requirement protease A3) is a trimeric PDZ bearing propapoptotic serine protease, which is involved in various diseases including cancer and pre-eclampsia. Proposed to be a tumor suppressor, its role as a potential therapeutic target is strongly advocated. Therefore, it becomes imperative to gain insights into its mechanism of action and regulation. Allostery is a well-known mechanism of catalytic activation for many HtrA3 homologs, which opens up avenues for manipulating enzyme functions for therapeutic intervention. In our study, through in silico and biochemical approaches, we have reported for the first time that HtrA3 shows allosteric behaviour. We identified a novel selective binding pocket, which triggers conformational reorientations through signal propagation to the distantly situated active-site pocket via the functionally important loop regions. Using molecular docking, simulation studies and biochemical studies we have identified the regulatory movements at and around the active site pocket. Our study is the first one to report a non-classical binding site for HtrA3, which is instrumental for formation of a catalytically efficient orthosteric pocket upon substrate binding. (C) 2019 Elsevier Inc. All rights reserved.

Cellular senescence has causative links with ageing and age-related diseases, however, it remains unclear if progeroid factors cause senescence in normal cells. Here, we show that depletion of CSB, a protein mutated in progeroid Cockayne syndrome (CS), is the earliest known trigger of p21-dependent replicative senescence. CSB depletion promotes overexpression of the HTRA3 protease resulting in mitochondrial impairments, which are causally linked to CS pathological phenotypes. The CSB promoter is downregulated by histone H3 hypoacetylation during DNA damage-response. Mechanistically, CSB binds to the p21 promoter thereby downregulating its transcription and blocking replicative senescence in a p53-independent manner. This activity of CSB is independent of its role in the repair of UV-induced DNA damage. HTRA3 accumulation and senescence are partially rescued upon reduction of oxidative/nitrosative stress. These findings establish a CSB/p21 axis that acts as a barrier to replicative senescence, and link a progeroid factor with the process of regular ageing in human.