Identification of a distal allosteric ligand binding pocket in HtrA3

Acharya, Saujanya; Dutta, Shubhankar; Chopra, Sucheta; Bose, Kakoli

doi:10.1016/j.bbrc.2019.07.005

Cited by 2 publications

(11 citation statements)

References 22 publications

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“…The absence of PDZ in this variant might lead to greater accessibility of the substrate to the active site; however, it also results in significant decrease in the catalytic efficiency, which might be due to a malformed oxyanion hole as observed previously in the case of its homolog HtrA2 [14]. Therefore, to understand the formation of the oxyanion hole upon β-casein binding, the β-casein peptide (AMVKLYC) [18] was docked with HtrA3 N-SPD at the allosteric site identified in our previous study [18]. Since some of the PDZ domain residues belonging to the allosteric pocket are missing in HtrA3 N-SPD, it resulted in a low docking score of −5.194 kcal/mole ( Figure 3B).…”

Section: Pdz Domain Is Indispensable For Optimal Catalytic Activationmentioning

confidence: 89%

“…HtrA3 N-SPD respectively (for mature HtrA3 V max = 1.95 ± 0.20 × 10 −9 M.s −1 , k cat = 0.56 ± 0.07 × 10 −3 s −1 and k cat /K 0.5 = 0.42 ± 0.03 × 10 3 M −1 .s −1 ) [18]. However, the K m was slightly decreased by 1.2-fold (for mature HtrA3 K 0.5 = 1.31 ± 0.06).…”

Section: Pdz Domain Is Indispensable For Optimal Catalytic Activationmentioning

confidence: 97%

“…It was previously determined that the activation of mature HtrA3 occurs allosterically [18]. To understand whether allosteric activation of HtrA3 requires any direct involvement of the C-terminal PDZ domain, we determined the enzymatic parameters for the PDZ-lacking variant (HtrA3 N-SPD) ( Table 2).…”

Section: Pdz Domain Is Indispensable For Optimal Catalytic Activationmentioning

confidence: 99%

“…For quantitative analysis, kinetic parameters for mature HtrA3 [18] and HtrA3 N-SPD were determined and our observations have been described in depth in the previous sections. HtrA3 SPD, HtrA3 F142D, HtrA3 F142A, HtrA3 F255D and HtrA3 SPD-PDZ were completely inactive hence no kinetic parameters could be obtained.…”

Section: Role Of Oligomerization Individual Domains and Critical Resmentioning

confidence: 99%

“…Briefly, literature available till date reports that the HtrA homologs assume a trimeric pyramidal architecture and exhibit similar mechanism of activation [11][12][13][14][15][16][17]. While HtrA-1, -2 and -4 have been reported to be allosterically activated [12,14,16], only one such study reports allostery in HtrA3 till date [18]. HtrA2, particularly, is allosterically regulated [12,14,16] through both the N-and C-terminal regions.…”

Section: Introductionmentioning

confidence: 99%

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A distinct concerted mechanism of structural dynamism defines activity of human serine protease HtrA3

Acharya

Dutta

Bose

2020

Biochemical Journal

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Human HtrA3 (high-temperature requirement protease A3) is a trimeric multitasking propapoptotic serine protease associated with critical cellular functions and pathogenicity. Implicated in diseases including cancer and pre-eclampsia, its role as a tumor suppressor and potential therapeutic target cannot be ignored. Therefore, elucidating its mode of activation and regulatory switch becomes indispensable towards modulating its functions with desired effects for disease intervention. Using computational, biochemical and biophysical tools, we delineated the role of all domains, their combinations and the critical phenylalanine residues in regulating HtrA3 activity, oligomerization and specificity. Our findings underline the crucial roles of the N-terminus as well as the PDZ domain in oligomerization and formation of a catalytically competent enzyme, thus providing new insights into its structure–function coordination. Our study also reports an intricate ligand-induced allosteric switch, which redefines the existing hypothesis of HtrA3 activation besides opening up avenues for modulating protease activity favorably through suitable effector molecules.

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Section: Pdz Domain Is Indispensable For Optimal Catalytic Activationmentioning

confidence: 89%

Section: Pdz Domain Is Indispensable For Optimal Catalytic Activationmentioning

confidence: 97%

Section: Pdz Domain Is Indispensable For Optimal Catalytic Activationmentioning

confidence: 99%

Section: Role Of Oligomerization Individual Domains and Critical Resmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A distinct concerted mechanism of structural dynamism defines activity of human serine protease HtrA3

Acharya

Dutta

Bose

2020

Biochemical Journal

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Hsa-miR-494-3p attenuates gene HtrA3 transcription to increase inflammatory response in hypoxia/reoxygenation HK2 Cells

Gong

Shen

Sheng

et al. 2021

Sci Rep

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The occurrence of cardiac surgery-associated acute kidney injury (CSA-AKI) increases hospital stay and mortality. MicroRNAs has a crucial role in AKI. This objective of the current study is to explore the function of hsa-miR-494-3p in inflammatory response in human kidney tubular epithelial (HK2) cells with hypoxia/reoxygenation. According to KDIGO standard, patients after cardiac surgery with cardiopulmonary bypass were divided into two groups: AKI (n = 10) and non-AKI patients (n = 8). HK2 were raised in the normal and hypoxia/reoxygenation circumstances and mainly treated by overexpression ofmiR-494-3p and HtrA3. The relationship between miR-494-3p and HtrA3 was determined by dual-luciferase reporter assay. Our result showed that Hsa-miR-494-3p was elevated in the serum of patients with CSA-AKI, and also induced in hypoxic reoxygenated HK2 cells. Hsa-miR-494-3p also increased a hypoxia-reoxygenation induced inflammatory response in HK2 cells. Moreover, as a target gene of miR-494-3p, overexpression of HtrA3 downregulated the hypoxia-reoxygenation induced inflammatory response in HK2 cells. Overexpression of hsa-miR-494-3p-induced inflammatory response was inhibited by overexpression of HtrA3. Collectively, we identified that hsa-miR-494-3p, a miRNA induced in both circulation of AKI patients and hypoxia-reoxygenation-treated HK2 cells, enhanced renal inflammation by targeting HtrA3, which may suggest a possible role as a new therapeutic target for CSA-AKI.

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Identification of a distal allosteric ligand binding pocket in HtrA3

Cited by 2 publications

References 22 publications

A distinct concerted mechanism of structural dynamism defines activity of human serine protease HtrA3

A distinct concerted mechanism of structural dynamism defines activity of human serine protease HtrA3

Hsa-miR-494-3p attenuates gene HtrA3 transcription to increase inflammatory response in hypoxia/reoxygenation HK2 Cells

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