Human plasma actin‐depolymerizing factor

Chaponnier, Christine; Patebex, Patricia; Gabbiani, Giulio

doi:10.1111/j.1432-1033.1985.tb08649.x

Cited by 33 publications

(15 citation statements)

References 48 publications

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“…The presence of multiple gelsolin bands in electrophoresis under native conditions is not unexpected, because gelsolin is resolved into 3 components by isoelectric focussing in urea . The markedly different mobility of the mutant gelsolin at pH 8.6 cannot be explained by the loss of a single negative charge in a protein with an isoelectric point about 6.2 [19,20,22], containing 98 acidic and 99 basic residues. The mobility difference most likely reflects the electrophoretic behaviour of the proteolytically cleaved mutant gelsolin.…”

Section: Discussionmentioning

confidence: 99%

Variant plasma gelsolin responsible for familial amyloidosis (Finnish type) has defective actin severing activity

et al. 1993

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Familial amyloidosis, Finnish type is caused by a single base mutation in gelsolin, an actin filament severing and capping protein that is present in most tissues and in blood plasma. The mutation replaces aspartic acid with asparagine at residue 187 of the plasma sequence. This renders the gelsolin susceptible to proteolysis as a consequence of which amyloid protein is formed. Here it is shown that the mutant protein in plasma from a patient homozygous for this mutation lacks both actin severing and nucleating activities. Evidence is presented that the cleaved mutant gelsolin has dissociated under non‐denaturing conditions and that the resultant 65,000 and 55,000 M r C‐terminal fragments aggregate.

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Section: Discussionmentioning

confidence: 99%

Variant plasma gelsolin responsible for familial amyloidosis (Finnish type) has defective actin severing activity

et al. 1993

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“…Preparation ofplu.sma gelsolin Plasma gelsolin was prepared from swine plasma according to the method previously developed [2] with a slight modification [31]. All operations were performed at 4°C or on ice.…”

Section: Methodsmentioning

confidence: 99%

Plasma-gelsolin-binding sites on the actin sequence

1987

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Plasma gelsolin was cross-linked to fluorescently labeled actin in order to identify plasma-gelsolin-binding sites on the primary sequence of actin. Plasma gelsolin can be cross-linked to actin with l-ethyl-3- [3-(dimethylamino)propyl]carbodiimide (EDC), resulting in the formation of 1 : 1 and 1 : 2 plasma gelsolin: actin cross-linked complexes with apparent molecular masses of 130 kDa and 180 kDa respectively. The cross-linked complexes were isolated separately, partially digested with cyanogen bromide and subjected to sodium dodecyl sulfate gel electrophoresis to analyze fluorescent fragments. The electrophoretic pattern showed that fluorescent CNBr fragments obtained from a free actin molecule were all found in those obtained from both the complexes. Since the fluorescent probe was attached to an actin molecule through the penultimate cysteine residue (Cys-374), the agreement in the fluorescent patterns indicated that the NH2-terminal CNBr fragments of both the actin molecules were involved in cross-linking with plasma gelsolin. This was also suggested by hydroxylamine cleavage of the two complexes as the cleavage gave the fluorescent 41-kDa fragment which could not be produced unless plasma gelsolin was cross-linked at the NH,-tenninal segment comprising 12 amino acids. Since EDC cross-links an amino group with a carboxyl group only when they are direct contact, the characteristic acidic amino acid residues at the NH2 terminus of actin are suggested to participate in binding plasma gelsolin.As one of the major components of the cytoskeleton, actin plays important roles in cell motility and its spatial organization. In non-muscle cells, actin plays these roles through the process of polymerization and depolymerization, which appears to be regulated by various actin-binding proteins [l]. Plasma gelsolin belongs to a group of the actinbinding proteins that bind to the fast growing (or 'barbed') end of actin filaments [2-41. Its functional and structural similarities to cytoplasmic gelsolin have been demonstrated [5]. Plasma gelsolin is also called actin-depolymerizing factor [6] or brevin [7]. The effect of plasma gelsolin on actin is Ca2 N-[7-(dimethylamino)-4-methyl-3-coumarinyl]maleimide. myosin S-1 binding sites on the primary sequence of actin [19]. He used fluorescent actin labeled selectively at the penultimate amino acid residue (Cys-374) for cross-linking with S-1, digested the isolated cross-linked actin-S-1 complex by CNBr, and subsequently analyzed the fluorescent products by one-dimensional peptide mapping to identify the binding sites for myosin on the actin sequence. The same cross-linking reagent was successfully used for identifying the binding sites of various types of actin-binding proteins showed that plasma gelsolin and actin were crosslinked by EDC to form 1 : 1 and 1 : 2 plasma gelsolin: actin cross-linked complexes. She indicated that the extent of crosslinking was much reduced at low calcium concentration [21]. The formation of 1 : 1 and 1 : 2 cross-linked complexes was also demonstra...

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“…The uncomplexed plasma gelsolin would then be free to sever actin filaments. These filaments might be released into the extracellular space, or found within dying cells where gelsolin can mediate the disassembly of the cytoskeleton (37). Higher levels of DBP (6-10 MM) as compared with plasma gelsolin (1)(2)(3) uM) may be present to provide a high level of monomerbinding protein, thereby ensuring the presence of a maximal amount of free plasma gelsolin, which is capable of severing filaments.…”

Section: Discussionmentioning

confidence: 99%

Role of plasma gelsolin and the vitamin D-binding protein in clearing actin from the circulation.

Lind¹,

Smith²,

Janmey³

et al. 1986

J. Clin. Invest.

152

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We determined the plasma kinetics of both actin and complexes of actin with the two high affinity actin-binding proteins of plasma, gelsolin, and vitamin D-binding protein (DBP). Actin is cleared rapidly from the plasma by the liver (half-disappearance time, 0.5 h). Using radiolabeled actin-binding proteins, we found that actin accelerated the clearance of both plasma gelsolin and the vitamin D-binding protein.In separate experiments we found that DBP-actin complexes were cleared more quickly than gelsolin-actin complexes, at a rate comparable to the clearance of actin from the blood. A low affinity interaction (dissociation constant, 2.9 X 10-M) between actin and fibronectin was found, suggesting that little actin will bind to fibronectin in plasma. We conclude that while plasma gelsolin and DBP may both clear actin from the circulation, DBP appears to play a more important role. By so doing, DBP may conserve the filament-severing activity of plasma gelsolin.

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Human plasma actin‐depolymerizing factor

Cited by 33 publications

References 48 publications

Variant plasma gelsolin responsible for familial amyloidosis (Finnish type) has defective actin severing activity

Variant plasma gelsolin responsible for familial amyloidosis (Finnish type) has defective actin severing activity

Plasma-gelsolin-binding sites on the actin sequence

Role of plasma gelsolin and the vitamin D-binding protein in clearing actin from the circulation.

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