Solution structure and dynamics of ADF from Toxoplasma gondii

Rahul Yadav, Prem Prakash Pathak, Vaibhav Kumar Shukla, Anupam Jain, Shubhra Srivastava, Sarita Tripathi, S. V.S.R. Krishna Pulavarti, Simren Mehta, L. David Sibley, Ashish Arora

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

Toxoplasma gondii ADF (TgADF) belongs to a functional subtype characterized by strong G-actin sequestering activity and low F-actin severing activity. Among the characterized ADF/cofilin proteins, TgADF has the shortest length and is missing a C-terminal helix implicated in F-actin binding. In order to understand its characteristic properties, we have determined the solution structure of TgADF and studied its backbone dynamics from 15N-relaxation measurements. TgADF has conserved ADF/cofilin fold consisting of a central mixed β-sheet comprised of six β-strands that are partially surrounded by three α-helices and a C-terminal helical turn. The high G-actin sequestering activity of TgADF relies on highly structurally and dynamically optimized interactions between G-actin and G-actin binding surface of TgADF. The equilibrium dissociation constant for TgADF and rabbit muscle G-actin was 23.81nM, as measured by ITC, which reflects very strong affinity of TgADF and G-actin interactions. The F-actin binding site of TgADF is partially formed, with a shortened F-loop that does not project out of the ellipsoid structure and a C-terminal helical turn in place of the C-terminal helix α4. Yet, it is more rigid than the F-actin binding site of Leishmania donovani cofilin. Experimental observations and structural features do not support the interaction of PIP2 with TgADF, and PIP2 does not affect the interaction of TgADF with G-actin. Overall, this study suggests that conformational flexibility of G-actin binding sites enhances the affinity of TgADF for G-actin, while conformational rigidity of F-actin binding sites of conventional ADF/cofilins is necessary for stable binding to F-actin.

Original languageEnglish
Pages (from-to)97-111
Number of pages15
JournalJournal of Structural Biology
Volume176
Issue number1
DOIs
StatePublished - Oct 1 2011

Keywords

  • G-actin
  • NMR
  • PIP2
  • Protein dynamics
  • TgADF
  • Toxoplasma gondii

Fingerprint Dive into the research topics of 'Solution structure and dynamics of ADF from Toxoplasma gondii'. Together they form a unique fingerprint.

  • Cite this

    Yadav, R., Pathak, P. P., Shukla, V. K., Jain, A., Srivastava, S., Tripathi, S., Krishna Pulavarti, S. V. S. R., Mehta, S., David Sibley, L., & Arora, A. (2011). Solution structure and dynamics of ADF from Toxoplasma gondii. Journal of Structural Biology, 176(1), 97-111. https://doi.org/10.1016/j.jsb.2011.07.011