Cysteine 81 is critical for the interaction of s100a4 and myosin-IIA

Natalia Dulianinova, Karen M. Hite, Wendy D. Zencheck, Dominic A. Scudiero, Steven C. Almo, Robert H. Shoemaker, Anne R. Bresnick

Research output: Contribution to journalArticle

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Abstract

Overexpression of S100A4, a member of the S100 family of Ca 2+-binding proteins, is associated with a number of human pathologies, including fibrosis, inflammatory disorders, and metastatic disease. The identification of small molecules that disrupt S100A4/target interactions provides a mechanism for inhibiting S100A4-mediated cellular activities and their associated pathologies. Using an anisotropy assay that monitors the Ca 2+-dependent binding of myosin-IIA to S100A4, NSC 95397 was identified as an inhibitor that disrupts the S100A4/myosin-IIA interaction and inhibits S100A4-mediated depolymerization of myosin-IIA filaments. Mass spectrometry demonstrated that NSC 95397 forms covalent adducts with Cys81 and Cys86, which are located in the canonical target binding cleft. Mutagenesis studies showed that covalent modification of just Cys81 is sufficient to inhibit S100A4 function with respect to myosin-IIA binding and depolymerization. Remarkably, substitution of Cys81 with serine or alanine significantly impaired the ability of S100A4 to promote myosin-IIA filament disassembly. As reversible covalent cysteine modifications have been observed for several S100 proteins, we propose that modification of Cys81 may provide an additional regulatory mechanism for mediating the binding of S100A4 to myosin-IIA.

Original languageEnglish (US)
Pages (from-to)7218-7227
Number of pages10
JournalBiochemistry
Volume50
Issue number33
DOIs
StatePublished - Aug 23 2011

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Nonmuscle Myosin Type IIA
Cysteine
2,3-bis(2-hydroxyethylsulfanyl)-(1,4)naphthoquinone
Depolymerization
Pathology
Mutagenesis
Aptitude
S100 Proteins
Anisotropy
Alanine
Serine
Mass spectrometry
Assays
Mass Spectrometry
Fibrosis
Substitution reactions
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cysteine 81 is critical for the interaction of s100a4 and myosin-IIA. / Dulianinova, Natalia; Hite, Karen M.; Zencheck, Wendy D.; Scudiero, Dominic A.; Almo, Steven C.; Shoemaker, Robert H.; Bresnick, Anne R.

In: Biochemistry, Vol. 50, No. 33, 23.08.2011, p. 7218-7227.

Research output: Contribution to journalArticle

Dulianinova N, Hite KM, Zencheck WD, Scudiero DA, Almo SC, Shoemaker RH et al. Cysteine 81 is critical for the interaction of s100a4 and myosin-IIA. Biochemistry. 2011 Aug 23;50(33):7218-7227. https://doi.org/10.1021/bi200853y
Dulianinova, Natalia ; Hite, Karen M. ; Zencheck, Wendy D. ; Scudiero, Dominic A. ; Almo, Steven C. ; Shoemaker, Robert H. ; Bresnick, Anne R. / Cysteine 81 is critical for the interaction of s100a4 and myosin-IIA. In: Biochemistry. 2011 ; Vol. 50, No. 33. pp. 7218-7227.
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