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Title: Structural and functional analysis of human HtrA3 protease and its subdomains

Journal Article · · PLoS ONE
 [1];  [2];  [1];  [1];  [1];  [1];  [3];  [1];  [2];  [1];  [4]
  1. Univ. of Gdansk (Poland). Dept. of Biochemistry.
  2. Argonne National Lab., Argonne, IL (United States). Midwest Center for Structural Genomics and Structural Biology Center.
  3. Univ. of Gdansk (Poland). Dept. of Molecular and Cellular Biology; Medical Univ. of Gdansk (Poland)
  4. Centro Nacional de Biotecnologia - CSIC (Spain)
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Human HtrA3 protease, which induces mitochondria-mediated apoptosis, can be a tumor suppressor and a potential therapeutic target in the treatment of cancer. However, there is little information about its structure and biochemical properties. HtrA3 is composed of an N-terminal domain not required for proteolytic activity, a central serine protease domain and a C-terminal PDZ domain. HtrA3S, its short natural isoform, lacks the PDZ domain which is substituted by a stretch of 7 C-terminal amino acid residues, unique for this isoform. This paper presents the crystal structure of the HtrA3 protease domain together with the PDZ domain (ΔN-HtrA3), showing that the protein forms a trimer whose protease domains are similar to those of human HtrA1 and HtrA2. The ΔN-HtrA3 PDZ domains are placed in a position intermediate between that in the flat saucer-like HtrA1 SAXS structure and the compact pyramidal HtrA2 X-ray structure. The PDZ domain interacts closely with the LB loop of the protease domain in a way not found in other human HtrAs. ΔN-HtrA3 with the PDZ removed (ΔN-HtrA3-ΔPDZ) and an N-terminally truncated HtrA3S (ΔN-HtrA3S) were fully active at a wide range of temperatures and their substrate affinity was not impaired. This indicates that the PDZ domain is dispensable for HtrA3 activity. As determined by size exclusion chromatography, ΔN-HtrA3 formed stable trimers while both ΔN-HtrA3-ΔPDZ and ΔN-HtrA3S were monomeric. This suggests that the presence of the PDZ domain, unlike in HtrA1 and HtrA2, influences HtrA3 trimer formation. The unique C-terminal sequence of ΔN-HtrA3S appeared to have little effect on activity and oligomerization. Additionally, we examined the cleavage specificity of ΔN-HtrA3. Results reported in this paper provide new insights into the structure and function of ΔN-HtrA3, which seems to have a unique combination of features among human HtrA proteases.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institute of Health; Polish National Science Centre (NCN)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1212403
Journal Information:
PLoS ONE, Vol. 10, Issue 6; ISSN 1932-6203
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 32 works
Citation information provided by
Web of Science

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Cited By (9)

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A distinct concerted mechanism of structural dynamism defines activity of human serine protease HtrA3 journal January 2020
Distinct 3D Architecture and Dynamics of the Human HtrA2(Omi) Protease and Its Mutated Variants journal August 2016
A Cell Proliferation and Inflammatory Signature is Induced by Lawsonia intracellularis Infection in Swine journal August 2018
Role of HTRA1 in bone formation and regeneration: In vitro and in vivo evaluation journal July 2017
Integration of genomic, transcriptomic and functional profiles of aggressive osteosarcomas across multiple species journal July 2017
High temperature requirement A3 (HTRA3) expression predicts postoperative recurrence and survival in patients with non-small-cell lung cancer journal May 2016
Properties of the HtrA Protease From Bacterium Helicobacter pylori Whose Activity Is Indispensable for Growth Under Stress Conditions journal May 2019
HtrA4 Protease Promotes Chemotherapeutic-Dependent Cancer Cell Death journal September 2019

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59 BASIC BIOLOGICAL SCIENCES
proteases
crystal structure
protein structure
peptides
oligomers
serine proteases
size-exclusion chromatography
X-ray crystallography