Differential Binding of Myosin Subfragment One Species to Immobilized ADP, and Actin: the Influence of the Alkali Light Chains

Winstanley, Monica A.; Small, David A.; Trayer, Ian P.

doi:10.1111/j.1432-1033.1979.tb13204.x

Cited by 24 publications

(5 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One plausible explanation for this is that S1 binding to the adjacent actin monomer displaces the N-terminal part of the A1 light chain from its interaction with the lower monomer. This is reasonable, considering the relative strengths and salt sensitivities of the S1 heavy chain and light chain binding to actin (Winstanley et al, 1979).…”

Section: Stoichiometry Of Myosin Binding To Actin Filamentssupporting

confidence: 52%

Interaction of Myosin with F-Actin: Time-Dependent Changes at the Interface Are Not Slow

Dijk

Céline

Barman³

et al. 2000

Biophysical Journal

View full text Add to dashboard Cite

The kinetics of formation of the actin-myosin complex have been reinvestigated on the minute and second time scales in sedimentation and chemical cross-linking experiments. With the sedimentation method, we found that the binding of the skeletal muscle myosin motor domain (S1) to actin filament always saturates at one S1 bound to one actin monomer (or two S1 per actin dimer), whether S1 was added slowly (17 min between additions) or rapidly (10 s between additions) to an excess of F-actin. The carbodiimide (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, EDC)-induced cross-linking of the actin-S1 complex was performed on the subsecond time scale by a new approach that combines a two-step cross-linking protocol with the rapid flow-quench technique. The results showed that the time courses of S1 cross-linking to either of the two actin monomers are identical: they are not dependent on the actin/S1 ratio in the 0.3-20-s time range. The overall data rule out a mechanism by which myosin rolls from one to the other actin monomer on the second or minute time scales. Rather, they suggest that more subtle changes occur at the actomyosin interface during the ATP cycle.

show abstract

Section: Stoichiometry Of Myosin Binding To Actin Filamentssupporting

confidence: 52%

Interaction of Myosin with F-Actin: Time-Dependent Changes at the Interface Are Not Slow

Dijk

Céline

Barman³

et al. 2000

Biophysical Journal

View full text Add to dashboard Cite

show abstract

“…The nucleotide-mediated dissociation of the acto-30-kDa system is observable not only with the 30-kDa peptide associated with both Al and A2 light chains but also with the 30-kDa-A2 complex (result not shown). Thus, in spite of its ability to interact with actin (Prince et al, 1981;Sutoh, 1982) and under some conditions with ATP (Winstanley et al, 1979;Burke et al, 1981), the Al light chain does not seem to contribute significantly to this process. The association of the 20-kDa peptide with actin was unaffected by any of the phosphorylated ligands tested.…”

Section: Resultsmentioning

confidence: 99%

Identification of polyphosphate recognition sites communicating with the actin sites on the skeletal myosin subfragment-1 heavy chain

Chaussepied¹,

Mornet²,

Kassab³

1986

Biochemistry

View full text Add to dashboard Cite

Using the thrombin-cut [68-30 kilodalton (kDa)] myosin subfragment 1 (S-1) whose heavy chain has been selectively split within the central 50-kDa region, at Lys-560, with concomitant specific alterations of the ATPase and actin binding properties [Chaussepied, P., Mornet, D., Audemard, E., Derancourt, J., & Kassab, R. (1986) Biochemistry 25, 1134-1140; Chaussepied, P., Mornet, D., Barman, T., Travers, F., & Kassab, R. (1986) Biochemistry 25, 1141-1149], we have isolated and renatured the COOH-terminal 30-kDa fragment associated with the alkali light chains by the procedure recently described [Chaussepied, P., Mornet, D., Audemard, E., Kassab, R., Goodearl, J., Levine, B., & Trayer, I. P. (1986) Biochemistry 25, 4540-4547]. The 30-kDa peptide preparation was found to exhibit a crucial feature of the native S-1; namely, it interacts with F-actin in an adenosine 5'-triphosphate (ATP)-dependent manner. Studies by ultracentrifugation, turbidity measurements, and chemical cross-linking experiments showed that the acto-30-kDa peptide complex was dissociated almost completely by the gamma-phosphoryl group containing ligands ATP, 5'-adenylyl imidodiphosphate, and pyrophosphate, to a lesser extent by ADP, and not at all by AMP and inorganic phosphate. The maximal dissociating effect is operating with the thrombic 30-kDa entity, whereas the 22-kDa fragment produced by staphylococcal protease is only slightly dissociated. In contrast, the tryptic 20-kDa fragment binds irreversibly to actin.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

“…Actin-activated MgATPase assays were carried out by the discontinuous assay procedure [33], keeping the ionic strength low in order to magnify the difference between S1(A1)-like and S1(A2)-like kinetics. All assays were carried out in buffer containing 5 mM triethanolamine/HCl, pH 7.5 at 25°C.…”

Section: Methodsmentioning

confidence: 99%

The N‐terminus of A1‐type myosin essential light chains binds actin and modulates myosin motor function

Timson

Trayer

1998

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

There are two isoforms (A1 and A2) of the myosin essential light chain (ELC) and consequently two isoenzymes of myosin subfragment 1 (S1), S1(A1) and S1(A2). The two isoenzymes differ in their kinetic properties with S1(A1) having a lower apparent K m for actin and a slower turnover of MgATP (k cat ) than S1(A2). The two forms of the ELC differ only at their N-termini where A1 has an additional 40-odd amino acids that are not present in A2.The human atrial ELC (an A1-type ELC) was overexpressed in Escherichia coli and purified by ammonium sulphate fractionation and ion-exchange chromatography. The recombinant ELC had actinactivated MgATPase kinetics similar to those for rabbit skeletal S1(A1) under the same conditions. Deletion of the first 45 amino acid residues resulted in an ELC similar to the rabbit skeletal A2 isoform and, when hybridised into S1, in S1(A2)-like kinetic properties. Results obtained with an ELC mutant that lacks the first 11 residues were intermediate between these two extremes but tending towards the S1(A2)-like phenotype.The wild-type ELC (both hybridised into S1 or free in solution) could be cross-linked to F-actin, whereas the deletion mutant lacking the first 45 amino acids could not. The deletion mutant lacking the first 11 amino acids cross-linked only poorly under the same conditions, consistent with the MgATPase data. We therefore conclude that these N-terminal eleven amino acids predominantly encode an actinbinding site which modulates the kinetics of the myosin motor. Furthermore, while free A1-type ELC cross-linked to both polymeric F-actin and the monomeric G-actin:DNase-I complex, the same ELC in S1(A1) could only cross-link to F-actin. This suggests that the light chain binds to a different actin monomer than the heavy chain.

show abstract

Differential Binding of Myosin Subfragment One Species to Immobilized ADP, and Actin: the Influence of the Alkali Light Chains

Cited by 24 publications

References 24 publications

Interaction of Myosin with F-Actin: Time-Dependent Changes at the Interface Are Not Slow

Interaction of Myosin with F-Actin: Time-Dependent Changes at the Interface Are Not Slow

Identification of polyphosphate recognition sites communicating with the actin sites on the skeletal myosin subfragment-1 heavy chain

The N‐terminus of A1‐type myosin essential light chains binds actin and modulates myosin motor function

Contact Info

Product

Resources

About