Structural and biochemical analyses of the tetrameric carboxypeptidase S9Cfn from Fusobacterium nucleatum. Issue 12 (18th November 2021)

• Record Type: Journal Article
• Authors: Wang, Xin; Cheng, Meng-Ting; Chen, Zhi-Peng; Jiang, Yong-Liang; Ge, Yu-Shu; Xia, Rong; Hou, Wen-Tao
• Is Part Of: Acta crystallographica. Volume 77:Issue 12(2021)
• Page Start: 1554
• Page End: 1563
• Publication Date: 2021-11-18
• Subjects: carboxypeptidases -- serine peptidases -- Fusobacterium nucleatum -- crystallography -- oral disease; X-ray crystallography -- Periodicals; Crystallography -- Periodicals; Molecular biology -- Periodicals; Molecular structure -- Periodicals; Biomolecules -- Structure -- Periodicals; Cytology -- Periodicals; Biomolecules -- Structure; Crystallography; Cytology; Molecular biology; Molecular structure; X-ray crystallography; Periodicals; 548
• DOI: 10.1107/S2059798321010810
• Languages: English
• ISSNs: 2059-7983

Abstract

Abstract : Extensive biochemical assays revealed that a putative peptidase S9Cfn from Fusobacterium nucleatum is a carboxypeptidase rather than an aminopeptidase as previously noted. Combined with the 2.6 Å tetramer structure of S9Cfn, key residues for substrate binding and catalysis, as well as the underlying mechanism of the catalytic cycle was revealed for S9C peptidase family. Abstract : As one of the most abundant bacteria in the human oral cavity, Fusobacterium nucleatum is closely involved in various oral diseases and is also a risk factor for other diseases. The peptidases of F. nucleatum can digest exogenous peptides into amino acids to satisfy its nutrient requirements. Here, a putative F. nucleatum peptidase, termed S9Cfn, which belongs to the S9C peptidase family was identified. Enzymatic activity assays combined with mass‐spectrometric analysis revealed that S9Cfn is a carboxypeptidase, but not an aminopeptidase as previously annotated. The crystal structure of the S9Cfn tetramer was solved at 2.6 Å resolution and was found to contain a pair of oligomeric pores in the center. Structural analysis, together with site‐directed mutagenesis and enzymatic activity assays, revealed a substrate‐entrance tunnel that extends from each oligomeric pore to the catalytic triad, adjacent to which three conserved arginine residues are responsible for substrate binding. Moreover, comparison with other S9 peptidase structures indicated drastic conformational changes of the oligomeric pores during the catalytic cycle. Together, these findings increase the knowledge of this unique type of tetrameric carboxypeptidase and provide insight into the homeostatic control of microbiota in the human oral cavity.