Mapping the functional surface of domain 2 in the gelsolin superfamily

Yoram A. Puius, Elena V. Fedorov, Ludwig Eichinger, Michael Schleicher, Steven C. Almo

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The crystal structure of the F-actin binding domain 2 of severin, the gelsolin homologue from Dictyostelium discoideum, has been determined by multiple isomorphous replacement and refined to 1.75 Å resolution. The structure reveals an α-helix-β-sheet sandwich similar to the domains of gelsolin and villin, and contains two cation-binding sites, as observed in other domain 1 and domain 2 homologues. Comparison of the structures of several gelsolin family domains has identified residues that may mediate F- actin binding in gelsolin domain 2 homologues. To assess the involvement of these residues in F-actin binding, three mutants of human gelsolin domain 2 were assayed for F-actin binding activity and thermodynamic stability. Two of the mutants, RRV168AAA and RLK210AAA, demonstrated a lowered affinity for F- actin, indicating a role for those residues in filament binding. Using both structural and biochemical data, we have constructed a model of the gelsolin domain 1-domain 2-F-actin complex. This model highlights a number of interactions that may serve as positive and negative determinants of filament end- and side-binding.

Original languageEnglish (US)
Pages (from-to)5322-5331
Number of pages10
JournalBiochemistry
Volume39
Issue number18
DOIs
StatePublished - May 9 2000

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Gelsolin
Actins
Dictyostelium
Thermodynamics
Cations
Thermodynamic stability
Crystal structure
Binding Sites

ASJC Scopus subject areas

  • Biochemistry

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Mapping the functional surface of domain 2 in the gelsolin superfamily. / Puius, Yoram A.; Fedorov, Elena V.; Eichinger, Ludwig; Schleicher, Michael; Almo, Steven C.

In: Biochemistry, Vol. 39, No. 18, 09.05.2000, p. 5322-5331.

Research output: Contribution to journalArticle

Puius, Yoram A. ; Fedorov, Elena V. ; Eichinger, Ludwig ; Schleicher, Michael ; Almo, Steven C. / Mapping the functional surface of domain 2 in the gelsolin superfamily. In: Biochemistry. 2000 ; Vol. 39, No. 18. pp. 5322-5331.
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AU - Almo, Steven C.

PY - 2000/5/9

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