The amino‐terminal fragment of gelsolin is cross‐linked to Cys‐374 of actin in the EGTA‐resistant actin‐gelsolin complex

Doi, Yukio; Kanatani, Yumiko; Kim, Fimi

doi:10.1016/0014-5793(92)80218-6

Cited by 5 publications

(2 citation statements)

References 33 publications

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“…An attempt to estimate accurate binding constants using lower concentrations of protein gave inconsistent results probably due to a high tendency toward denaturation of gelsolin at low concentrations, as observed previously (Weeds et al, 1986;Doi et al, 1991b). Together with our previous observations (Doi et al, 1987(Doi et al, , 1991a(Doi et al, , 1992) and those of others (Sutoh & Yin, 1989), we can specify a geometrical relationship of two actin molecules associating with gelsolin (Figure 9). In the EGTA-resistant actin, Cys-374 is 9-10 A apart from the amino terminal (residues 1-133) fragment of gelsolin and one of the first five acidic residues of the amino terminus is also in direct contact with the same amino terminal fragment of gelsolin.…”

Section: Discussionsupporting

confidence: 74%

Interaction of gelsolin with covalently cross-linked actin dimer

Doi

1992

Biochemistry

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One of the two actin molecules in the ternary actin-gelsolin complex was selectively cross-linked to gelsolin when benzophenonemaleimide-actin (BPM-actin) was used [Doi, Y., Banba, M., & Vertut-Doi (1991a) Biochemistry 30, 5769-5777]. Here, we examine the interaction between gelsolin and BPM-actin dimer in which BPM-actin is covalently conjugated to unlabeled actin by p-phenylenedimaleimide (pPDM). BPM-actin dimer having an apparent molecular mass of 115 kDa is photo-cross-linked to gelsolin (90 kDa) more effectively than BPM-actin monomer in the presence of Ca2+, forming a cross-linked actin dimer-gelsolin (1:1) complex with a molecular mass of 210 kDa. The tight direct association of the dimer to gelsolin is shown by the titration of gelsolin with the fluorescently labeled dimer and by the higher concentration of phosphatidylinositol 4,5-bisphosphate required to inhibit the formation of BPM-dimer complex with gelsolin than that of BPM-monomer complex. However, an attempt to cross-link the two actin molecules in the ternary actin-gelsolin (2:1) complex by pPDM fails. The results argue that the topography of the two actin molecules in the actin-gelsolin (2:1) complex is similar, but not identical, to that of the barbed end of an actin filament.

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Section: Discussionsupporting

confidence: 74%

Interaction of gelsolin with covalently cross-linked actin dimer

Doi

1992

Biochemistry

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“…Gelsolin modifies the conformation of the C‐terminus through direct interactions with C374 (Doi et al, 1992). Gelsolin binding leads to a shift in the C‐terminus and conformational changes propagating to the D‐loop, promoting filament nucleation (Hild et al, 2010; Khaitlina & Hinssen, 1997; Orlova et al, 1995).…”

Section: Actin Allostery and The C‐terminusmentioning

confidence: 99%

Actin's C‐terminus coordinates actin structural changes and functions

Steffensen

Dawson

2023

Cytoskeleton

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Actin is essential to eukaryotic cellular processes. Actin's C‐terminus appears to play a direct role in modulating actin's structure and properties, facilitating the binding and function of actin‐binding proteins (ABPs). The structural and functional characterization of filamentous actin's C‐terminus has been impeded by its inherent flexibility, as well as actin's resistance to crystallization for x‐ray diffraction and the historical resolution constraints associated with electron microscopy. Many biochemical studies have established that actin's C‐terminus must retain its flexibility and structural integrity to modulate actin's structure and functions. For example, C‐terminal structural changes are known to affect nucleotide binding and exchange, as well as propagate actin structural changes throughout extensive allosteric networks, facilitating the binding and function of ABPs. Advances in electron microscopy have resulted in high‐resolution structures of filamentous actin, providing insights into subtle structural changes that are mediated by actin's C‐terminus. Here, we review existing knowledge establishing the importance of actin's C‐terminus within actin structural changes and functions and discuss how modern structural characterization techniques provide the tools to understand the role of actin's C‐terminus in cellular processes.

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Conformational and functional studies of three gelsolin subdomain-1 synthetic peptides and their implication in actin polymerization

et al. 1997

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Gelsolin, a calcium and inositol phospholipid‐sensitive protein, regulates actin filament length. Its activity is complex (capping, severing, etc.) and is supported by several functional domains. The N‐terminal domain alone (S1), in particular, is able to impede actin polymerization. Our investigations were attempted to precise this inhibitory process by using synthetic peptides as models mimicking gelsolin S1 activity. Three peptides issued from S1 and located in gelsolin—actin interfaces were synthesized. The peptides (15–28, 42–55, and 96–114 sequences) were tested for their conformational and actin binding properties. Although the three peptides interact well with actin, only peptide 42–55 affects actin polymerization. A detailed kinetic study shows that the latter peptide essentially inhibits the nucleation step during actin polymerization. In conclusion, the present work shows that the binding of a synthetic peptide to a small sequence located outside the actin—actin interface is essential in the actin polymerization process. © 1997 John Wiley & Sons, Inc. Biopoly 41: 647–655, 1997

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The amino‐terminal fragment of gelsolin is cross‐linked to Cys‐374 of actin in the EGTA‐resistant actin‐gelsolin complex

Abstract: It has been shown thut the EGTA-resistant uctin. one of the two actin molcculcs ussociutcd to gclsolin. can bc prcdominuntly

Cited by 5 publications

References 33 publications

Interaction of gelsolin with covalently cross-linked actin dimer

Interaction of gelsolin with covalently cross-linked actin dimer

Actin's C‐terminus coordinates actin structural changes and functions

Conformational and functional studies of three gelsolin subdomain-1 synthetic peptides and their implication in actin polymerization

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