Ca2+ binding by domain 2 plays a critical role in the activation and stabilization of gelsolin

Shalini Nag, Qing Ma, Hui Wang, Sakesit Chumnarnsilpa, Lin Lee Wei, Mårten Larsson, Balakrishnan Kannan, Maria Hernandez-Valladares, Leslie D. Burtnick, Robert C. Robinson

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Gelsolin consists of six homologous domains (G1-G6), each containing a conserved Ca-binding site. Occupation of a subset of these sites enables gelsolin to sever and cap actin filaments in a Ca-dependent manner. Here, we present the structures of Ca-free human gelsolin and of Ca-bound human G1-G3 in a complex with actin. These structures closely resemble those determined previously for equine gelsolin. However, the G2 Ca-binding site is occupied in the human G1-G3/actin structure, whereas it is vacant in the equine version. In-depth comparison of the Ca-free and Ca-activated, actin-bound human gelsolin structures suggests G2 and G6 to be cooperative in binding Ca2+ and responsible for opening the G2-G6 latch to expose the F-actin-binding site on G2. Mutational analysis of the G2 and G6 Ca-binding sites demonstrates their interdependence in maintaining the compact structure in the absence of calcium. Examination of Ca binding by G2 in human G1-G3/actin reveals that the Ca 2+ locks the G2-G3 interface. Thermal denaturation studies of G2-G3 indicate that Ca binding stabilizes this fragment, driving it into the active conformation. The G2 Ca-binding site is mutated in gelsolin from familial amyloidosis (Finnish-type) patients. This disease initially proceeds through protease cleavage of G2, ultimately to produce a fragment that forms amyloid fibrils. The data presented here support a mechanism whereby the loss of Ca binding by G2 prolongs the lifetime of partially activated, intermediate conformations in which the protease cleavage site is exposed.

Original languageEnglish
Pages (from-to)13713-13718
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number33
DOIs
Publication statusPublished - Aug 18 2009
Externally publishedYes

Keywords

  • Actin
  • Calcium activated
  • Calcium dependent
  • TIRF

ASJC Scopus subject areas

  • General

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    Nag, S., Ma, Q., Wang, H., Chumnarnsilpa, S., Wei, L. L., Larsson, M., Kannan, B., Hernandez-Valladares, M., Burtnick, L. D., & Robinson, R. C. (2009). Ca2+ binding by domain 2 plays a critical role in the activation and stabilization of gelsolin. Proceedings of the National Academy of Sciences of the United States of America, 106(33), 13713-13718. https://doi.org/10.1073/pnas.0812374106