3TI9

Crystal structure of the basic protease BprB from the ovine footrot pathogen, Dichelobacter nodosus

3TI9 の概要

• エントリーDOI: 10.2210/pdb3ti9/pdb
• 関連するPDBエントリー: 3LPA 3TI7
• 分子名称: Serine protease, CALCIUM ION, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: hydrolase
• 由来する生物種: Dichelobacter nodosus
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 37347.60

構造登録者

Wong, W.,Whisstock, J.C.,Porter, C.J. (登録日: 2011-08-20, 公開日: 2011-10-19, 最終更新日: 2024-11-06)

主引用文献

Wong, W.,Wijeyewickrema, L.C.,Kennan, R.M.,Reeve, S.B.,Steer, D.L.,Reboul, C.,Smith, A.I.,Pike, R.N.,Rood, J.I.,Whisstock, J.C.,Porter, C.J.
S1 Pocket of a Bacterially Derived Subtilisin-like Protease Underpins Effective Tissue Destruction.
J.Biol.Chem., 286:42180-42187, 2011

PubMed Abstract: The ovine footrot pathogen, Dichelobacter nodosus, secretes three subtilisin-like proteases that play an important role in the pathogenesis of footrot through their ability to mediate tissue destruction. Virulent and benign strains of D. nodosus secrete the basic proteases BprV and BprB, respectively, with the catalytic domain of these enzymes having 96% sequence identity. At present, it is not known how sequence variation between these two putative virulence factors influences their respective biological activity. We have determined the high resolution crystal structures of BprV and BprB. These data reveal that that the S1 pocket of BprV is more hydrophobic but smaller than that of BprB. We show that BprV is more effective than BprB in degrading extracellular matrix components of the host tissue. Mutation of two residues around the S1 pocket of BprB to the equivalent residues in BprV dramatically enhanced its proteolytic activity against elastin substrates. Application of a novel approach for profiling substrate specificity, the Rapid Endopeptidase Profiling Library (REPLi) method, revealed that both enzymes prefer cleaving after hydrophobic residues (and in particular P1 leucine) but that BprV has more restricted primary substrate specificity than BprB. Furthermore, for P1 Leu-containing substrates we found that BprV is a significantly more efficient enzyme than BprB. Collectively, these data illuminate how subtle changes in D. nodosus proteases may significantly influence tissue destruction as part of the ovine footrot pathogenesis process.

PubMed: 21990366
DOI: 10.1074/jbc.M111.298711

実験手法

X-RAY DIFFRACTION (1.8 Å)