Structure of the S100A4/myosin-IIA complex

Udupi A. Ramagopal, Natalia Dulianinova, Kristen M. Varney, Paul T. Wilder, Sridevi Nallamsetty, Michael D. Brenowitz, David J. Weber, Steven C. Almo, Anne R. Bresnick

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: S100A4, a member of the S100 family of Ca§ssup§ 2+§esup§-binding proteins, modulates the motility of both non-transformed and cancer cells by regulating the localization and stability of cellular protrusions. Biochemical studies have demonstrated that S100A4 binds to the C-terminal end of the myosin-IIA heavy chain coiled-coil and disassembles myosin-IIA filaments; however, the mechanism by which S100A4 mediates myosin-IIA depolymerization is not well understood. Results: We determined the X-ray crystal structure of the S100A4Δ8C/MIIA§ssup§1908- 1923§esup§ peptide complex, which showed an asymmetric binding mode for the myosin-IIA peptide across the S100A4 dimer interface. This asymmetric binding mode was confirmed in NMR studies using a spin-labeled myosin-IIA peptide. In addition, our NMR data indicate that S100A4Δ8C binds the MIIA§ssup§1908-1923§esup§ peptide in an orientation very similar to that observed for wild-type S100A4. Studies of complex formation using a longer, dimeric myosin-IIA construct demonstrated that S100A4 binding dissociates the two myosin-IIA polypeptide chains to form a complex composed of one S100A4 dimer and a single myosin-IIA polypeptide chain. This interaction is mediated, in part, by the instability of the region of the myosin-IIA coiled-coil encompassing the S100A4 binding site. Conclusion: The structure of the S100A4/MIIA§ssup§1908-1923§esup§ peptide complex has revealed the overall architecture of this assembly and the detailed atomic interactions that mediate S100A4 binding to the myosin-IIA heavy chain. These structural studies support the idea that residues 1908-1923 of the myosin-IIA chain heavy represent a core sequence for the S100A4/myosin-IIA complex. In addition, biophysical studies suggest that structural fluctuations within the myosin-IIA coiled-coil may facilitate S100A4 docking onto a single myosin-IIA polypeptide chain.

Original languageEnglish (US)
Article number31
JournalBMC Structural Biology
Volume13
Issue number1
DOIs
StatePublished - Nov 20 2013

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Nonmuscle Myosin Type IIA
Peptides
Myosin Heavy Chains
Cell Surface Extensions

Keywords

  • Coiled-coil
  • Cytoskeleton
  • Myosin-II
  • NMR
  • S100A4
  • X-ray crystallography

ASJC Scopus subject areas

  • Structural Biology

Cite this

Structure of the S100A4/myosin-IIA complex. / Ramagopal, Udupi A.; Dulianinova, Natalia; Varney, Kristen M.; Wilder, Paul T.; Nallamsetty, Sridevi; Brenowitz, Michael D.; Weber, David J.; Almo, Steven C.; Bresnick, Anne R.

In: BMC Structural Biology, Vol. 13, No. 1, 31, 20.11.2013.

Research output: Contribution to journalArticle

Ramagopal, Udupi A. ; Dulianinova, Natalia ; Varney, Kristen M. ; Wilder, Paul T. ; Nallamsetty, Sridevi ; Brenowitz, Michael D. ; Weber, David J. ; Almo, Steven C. ; Bresnick, Anne R. / Structure of the S100A4/myosin-IIA complex. In: BMC Structural Biology. 2013 ; Vol. 13, No. 1.
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AU - Dulianinova, Natalia

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AU - Wilder, Paul T.

AU - Nallamsetty, Sridevi

AU - Brenowitz, Michael D.

AU - Weber, David J.

AU - Almo, Steven C.

AU - Bresnick, Anne R.

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N2 - Background: S100A4, a member of the S100 family of Ca§ssup§ 2+§esup§-binding proteins, modulates the motility of both non-transformed and cancer cells by regulating the localization and stability of cellular protrusions. Biochemical studies have demonstrated that S100A4 binds to the C-terminal end of the myosin-IIA heavy chain coiled-coil and disassembles myosin-IIA filaments; however, the mechanism by which S100A4 mediates myosin-IIA depolymerization is not well understood. Results: We determined the X-ray crystal structure of the S100A4Δ8C/MIIA§ssup§1908- 1923§esup§ peptide complex, which showed an asymmetric binding mode for the myosin-IIA peptide across the S100A4 dimer interface. This asymmetric binding mode was confirmed in NMR studies using a spin-labeled myosin-IIA peptide. In addition, our NMR data indicate that S100A4Δ8C binds the MIIA§ssup§1908-1923§esup§ peptide in an orientation very similar to that observed for wild-type S100A4. Studies of complex formation using a longer, dimeric myosin-IIA construct demonstrated that S100A4 binding dissociates the two myosin-IIA polypeptide chains to form a complex composed of one S100A4 dimer and a single myosin-IIA polypeptide chain. This interaction is mediated, in part, by the instability of the region of the myosin-IIA coiled-coil encompassing the S100A4 binding site. Conclusion: The structure of the S100A4/MIIA§ssup§1908-1923§esup§ peptide complex has revealed the overall architecture of this assembly and the detailed atomic interactions that mediate S100A4 binding to the myosin-IIA heavy chain. These structural studies support the idea that residues 1908-1923 of the myosin-IIA chain heavy represent a core sequence for the S100A4/myosin-IIA complex. In addition, biophysical studies suggest that structural fluctuations within the myosin-IIA coiled-coil may facilitate S100A4 docking onto a single myosin-IIA polypeptide chain.

AB - Background: S100A4, a member of the S100 family of Ca§ssup§ 2+§esup§-binding proteins, modulates the motility of both non-transformed and cancer cells by regulating the localization and stability of cellular protrusions. Biochemical studies have demonstrated that S100A4 binds to the C-terminal end of the myosin-IIA heavy chain coiled-coil and disassembles myosin-IIA filaments; however, the mechanism by which S100A4 mediates myosin-IIA depolymerization is not well understood. Results: We determined the X-ray crystal structure of the S100A4Δ8C/MIIA§ssup§1908- 1923§esup§ peptide complex, which showed an asymmetric binding mode for the myosin-IIA peptide across the S100A4 dimer interface. This asymmetric binding mode was confirmed in NMR studies using a spin-labeled myosin-IIA peptide. In addition, our NMR data indicate that S100A4Δ8C binds the MIIA§ssup§1908-1923§esup§ peptide in an orientation very similar to that observed for wild-type S100A4. Studies of complex formation using a longer, dimeric myosin-IIA construct demonstrated that S100A4 binding dissociates the two myosin-IIA polypeptide chains to form a complex composed of one S100A4 dimer and a single myosin-IIA polypeptide chain. This interaction is mediated, in part, by the instability of the region of the myosin-IIA coiled-coil encompassing the S100A4 binding site. Conclusion: The structure of the S100A4/MIIA§ssup§1908-1923§esup§ peptide complex has revealed the overall architecture of this assembly and the detailed atomic interactions that mediate S100A4 binding to the myosin-IIA heavy chain. These structural studies support the idea that residues 1908-1923 of the myosin-IIA chain heavy represent a core sequence for the S100A4/myosin-IIA complex. In addition, biophysical studies suggest that structural fluctuations within the myosin-IIA coiled-coil may facilitate S100A4 docking onto a single myosin-IIA polypeptide chain.

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KW - Cytoskeleton

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