Interactions of gelsolin and gelsolin-actin complexes with actin. Effects of calcium on actin nucleation, filament severing, and end blocking

Janmey, Paul A.; Chaponnier, Christine; Lind, Stuart E.; Zaner, Ken S.; Stossel, Thomas P.; Yin, Helen L.

doi:10.1021/bi00335a046

Cited by 195 publications

(151 citation statements)

References 33 publications

Supporting

Mentioning

136

Contrasting

Order By: Relevance

“…80% of the bound gelsolin was pelleted a second time in EGTA, compared to 94% in calcium. Janmey et al [33] have reported that actin filaments capped with macrophage gelsolin also showed incomplete dissociation of the gelsolin when calcium was removed. This is not surprising in view of the very tight association between the two proteins.…”

Section: Discussionmentioning

confidence: 99%

Binding of pig plasma gelsolin to F‐actin and partial fractionation into calcium‐dependent and calcium‐independent forms

Pope

Weeds

1986

European Journal of Biochemistry

View full text Add to dashboard Cite

The interaction of pig plasma gelsolin with F‐actin has been studied by a sedimentation assay using 125I‐gelsolin in a Beckman Airfuge. Over 90% of the gelsolin bound to F‐actin in 0.1 mM CaCl2 in experiments using 24 μM actin and 2–10 nM 125I‐gelsolin, but only 40–50% bound in 1 mM EGTA. Addition of more F‐actin to the EGTA supernantant does not sediment this gelsolin. Demonstration of this partial calcium sensitivity depends critically on the use of F‐actin that has been prepared in the absence of calcium ions. F‐actin prepared from G‐actin in calcium or pretreated with calcium, binds 125I‐gelsolin more completely in EGTA. This suggests that gelsolin activity is influenced by transient exposure of actin to calcium. Further evidence for partial calcium sensitivity in the interactions between gelsolin and F‐actin has been obtained by other methods, including viscometry and electron microscopy. The gelsolin present in the EGTA supernatant is complexed to G‐actin, predominantly as binary complexes. Very low concentrations of these complexes reduce the viscosity of F‐actin in calcium but not in EGTA. Whether this effect is due to severing activity, or capping with consequent depolymerization to establish the new critical concentration, is uncertain. The results suggest the presence of two types of gelsolin, one that requires micromolar concentrations of calcium for binding to F‐actin and one that does not. Both bind to G‐actin. Partial separation has been achieved using actin‐Sepharose. Pig plasma gelsolin is heterogeneous on isoelectric focussing gels in urea, but the two types of gelsolin separated on actin‐Sepharose do not correspond to specific isoelectric species.

show abstract

Section: Discussionmentioning

confidence: 99%

Binding of pig plasma gelsolin to F‐actin and partial fractionation into calcium‐dependent and calcium‐independent forms

Pope

Weeds

1986

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…The nucleating and severing activities are calcium dependent, but removal of calcium does not reverse all actin binding [3-51. For example, only one monomer dissociates from the ternary complex, leaving a binary complex containing trapped calcium, which cannot be dissociated without denaturation [3]. Similar properties have been reported for gelsolins from a variety of sources, including rabbit macrophage [6], human plasma [7] and pig stomach smooth muscle [8]. In contrast, pig plasma gelsolin has been shown to interact with actin both in the presence and absence of calcium [9-111.…”

Section: Introductionmentioning

confidence: 87%

“…Purification of cytoplasmic gelsolins has conventionally been carried out in the presence of EGTA (to prevent interaction with cellular actin) [6], while plasma gelsolin has been purified by affinity chromatography on actin-Sepharose in the presence of calcium [14] (blood plasma contains about 2 mM calcium). Cytoplasmic gelsolin-actin complexes have also been purified in the presence of calcium by affinity chromatography on DNase I-agarose [ 151, but subsequent separation of the complexes in 6 M urea was done in 1 mM EGTA [3].…”

Section: Introductionmentioning

confidence: 99%

Loss of calcium sensitivity of plasma gelsolin is associated with the presence of calcium ions during preparation

et al. 1989

View full text Add to dashboard Cite

Gelsolin is a calcium-dependent actin severing and capping protein. Calcium 'opens' the molecule to make actin binding sites accessible, but removal of calcium from the medium does not necessarily fully reverse this process. The calcium sensitivity of actin monomer binding and actin filament severing is here shown to vary considerably with the source of gelsolin and conditions of preparation. Plasma gelsolin undergoes irreversible loss of calcium sensitivity when prepared in the presence of calcium ions. This is not due solely to effects of bound calcium, because purified human plasma gelsolin expressed in E. coli and stored in calcium shows no comparable loss of calcium sensitivity when prepared or stored in calcium. These results suggest the presence of factors in plasma which, in the presence of calcium, promote an irreversible structural change in gelsolin resulting in permanent loss of calcium sensitivity.

show abstract

“…In vitro, villin functions as a bundling protein and does not sever actin filaments (anmey and Matsudaira, 1988;Petrucci et al, 1988); gCap39 binding to actin is reversible by Ca++ chelation and has limited ability to sever filaments (Southwick and DiNubile, 1986;Young et al, 1990;Yu et al, 1990). In contrast, gelsolin severs actin filaments, caps the barbed-end of filaments, and is not dissociated from actin by Ca++ chelation but only by exposure to PIP2 micelles (Janmey et al, 1985Janmey and Matsudaira, 1988). Because villin is not found in platelets or PMNs, because [Ca++]i is "tonically" elevated (for .5 min) (Sullivan et al, 1989) in FMLP activated PMN, and because ligand activation causes both Ca++ transient and PIP2 turnover (Pozzan et al, 1983;Cockcroft et al, 1985;Lapetina, 1990;Stephens et al, 1991), we investigated the role of gelsolin as a plausible regulator and creator of barbed-end nuclei in FMLP activated neutrophils.…”

Section: Ga Interactions In Basal Etf-pmnsmentioning

confidence: 99%

“…Interactions of gelsolin with actin in vitro are regulated by Ca++ and PIP2 (Yin and Stossel, 1979;Yin et al, 1981b;Janmey et al, 1985. In vitro micromolar Ca++ activates gelsolin's ability to sever filamentous (F) actin, cap the barbed (+) end of F-actin, and nucleate globular (G) actin polymerization at the pointed end of the filament.…”

Section: Introductionmentioning

confidence: 99%

Role of gelsolin interaction with actin in regulation and creation of actin nuclei in chemotactic peptide activated polymorphonuclear neutrophils.

Deaton

Guerrero

Howard

1992

MBoC

View full text Add to dashboard Cite

In vitro Ca++ activates gelsolin to sever F-actin and form a gelsolin-actin (GA) complex at the + end of F-actin that is not dissociated by ethylene glycol-bis(3-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) but is separated by EGTA + PIP/PIP2. The gelsolin blocks the + end on the actin filament, but the -end of the filament can still initiate actin polymerization. In thrombin activated platelets, evidence suggests that severing of F-actin by gelsolin increases GA complex, creates one -end actin nucleus and one cryptic + end actin nucleus per cut, and then dissociates to yield free + ends to nucleate rapid actin assembly. We examined the role of F-actin severing in creation and regulation of nuclei and polymerization in polymorphonuclear neutrophils (PMNs). At 2-s intervals after formyl peptide (FMLP) activation of endotoxin free (ETF) PMNs, change in GA complex was correlated with change in + end actin nuclei, -end actin nuclei, and F-actin content. GA complex was quantitated by electrophoretograms of proteins absorbed by antigelsolin from cells lysed in 10 mM EGTA, + end actin nuclei as cytochalasin (CD) sensitive and -end actin nuclei as CD insensitive increases in G-pyrenyl actin polymerization rates induced by the same PMNs, and F-actin content by NBDphallacidin binding to fixed cells. Thirty three percent of gelsolin was in GA complex in basal ETF PMNs; from 2-6 s, GA complexes dissociate (low = 15% at 10 s) and sequentially + end nuclei and F-actin content and then -end nuclei increase to a maximum at 10 s. At >40 s GA complex increase toward basal and + end nuclei and F-actin content returned toward basal. These kinetic data show gelsolin regulates availability of + end nuclei and actin polymerization in FMLP. However, absence of an initial increase in GA complex or -end nucleating activity shows FMLP activation does not cause gelsolin to sever F-or to bind G-actin to create cryptic + end nuclei in PMNs; the results suggest the + nucleus formation is gelsolin independent.

show abstract

Interactions of gelsolin and gelsolin-actin complexes with actin. Effects of calcium on actin nucleation, filament severing, and end blocking

Cited by 195 publications

References 33 publications

Binding of pig plasma gelsolin to F‐actin and partial fractionation into calcium‐dependent and calcium‐independent forms

Binding of pig plasma gelsolin to F‐actin and partial fractionation into calcium‐dependent and calcium‐independent forms

Loss of calcium sensitivity of plasma gelsolin is associated with the presence of calcium ions during preparation

Role of gelsolin interaction with actin in regulation and creation of actin nuclei in chemotactic peptide activated polymorphonuclear neutrophils.

Contact Info

Product

Resources

About