Structural analysis of inter-genus complexes of V-antigen and its regulator and their stabilization by divalent metal ions

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• 作者：Abhishek Basu ; Atanu Das ; Abhisek Mondal ; Saumen Datta
• 关键词：Inter ; genus complex ; Structural identity ; Elongated conformation ; Nanomolar binding affinity ; Polydispersity index ; Helix ; helix stabilization
• 刊名：European Biophysics Journal
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：45
• 期：2
• 页码：113-128
• 全文大小：1,808 KB
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• 作者单位：Abhishek Basu (1)
  Atanu Das (1)
  Abhisek Mondal (1)
  Saumen Datta (1)
  
  1. Structural Biology and Bioinformatics Division, CSIR-Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Kolkata, 700032, West Bengal, India
  
• 刊物类别：Physics and Astronomy
• 刊物主题：Physics
  Biophysics and Biomedical Physics
  Cell Biology
  Biochemistry
  Plant Physiology
  Animal Physiology
  Neurobiology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-1017

文摘

Gram-negative bacteria like Yersinia, Pseudomonas, and Aeromonas need type III secretion system (T3SS) for their pathogenicity. V-antigen and its regulator are essential for functioning of T3SS. There is significant functional conservation amongst V-antigen and its regulator belonging to the Ysc family. In this study, we have structurally characterized the inter-genus complexes of V-antigen and its regulator. ConSurf analysis demonstrates that V-antigens belonging to the Ysc family show high structural identity predominantly confined to the two long helical regions. The regulator of V-antigen shows high conservation in its first intramolecular coiled-coil domain, responsible for interaction with V-antigen. ∆LcrG<sub>(1–70)sub> localizes within the groove formed by long helices of LcrV, as observed in PcrV-∆PcrG<sub>(13–72)sub> interaction. Inter-genus complexes of LcrV-PcrG and PcrV-LcrG exhibited elongated conformation and 1:1 heterodimeric state like the native complex of PcrV-PcrG and LcrV-LcrG. Both native and inter-genus complexes showed rigid tertiary structure, solvent-exposed hydrophobic patches, and cooperative melting behavior with high melting temperature. LcrV-PcrG and PcrV-LcrG showed nanomolar affinity of interaction, identical to PcrV-PcrG interaction, but stronger than LcrV-LcrG interaction. Calcium (a secretion blocker of T3SS) propels all the complexes towards a highly monodisperse form. Calcium and magnesium increase the helicity of the native and inter-genus complexes, and causes helix–helix stabilization. Stabilization of helices leads to a slight increase in the melting temperature by 1.5–2.0 °C. However, calcium does not alter the affinity of interaction of V-antigen and its regulator, emphasizing the effect of divalent of cations at the structural level without any regulatory implications. Therefore, the structural conservation of these inter-genus complexes could be the basis for their functional complementation. Keywords Inter-genus complex Structural identity Elongated conformation Nanomolar binding affinity Polydispersity index Helix-helix stabilization