Crosslinking of the skeletal myosin subfragment-1 heavy chain to the N-terminal actin segment of residues 40-113

Bertrand, Raoul; Chaussepied, Patrick; Kassab, Ridha; Boyer, Mireille; Roustan, Claude; Benyamin, Yves

doi:10.1021/bi00415a050

Cited by 63 publications

(64 citation statements)

References 52 publications

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“…The strong cationic environment of this region involved in electrostatic actin-S-I interactions [11] might have been the determinant of the activity of the enzyme on S-1. The new nicked S-1 and myosin derivatives are useful for analyzing the functions of the intact 75 kDa domain [26,27].…”

Section: Localization Of the Arg-c Clip Site On The S-1 Heavy Chainmentioning

confidence: 99%

Selective cleavage at lysine of the 50 kDa‐20 kDa connector loop segment of skeletal myosin S‐1 by endoproteinase Arg‐C

1989

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The reaction of endoproteinase Arg-C on the skeletal myosin head heavy chain was investigated through characterization of peptides and amino acid sequence analysis. The protease splits exclusively the 50 kDa-20 kDa junction at the lysine cluster spanning residues 639~i41 and does not affect any other protease-sensitive region of the entire myosin heavy chain. The sensitivity of the cleavage to actin and nucleotide binding makes this protease a very specific conformational probe of S-1. The nicked S-1 derivative, containing an intact NH2-terminal 75 kDa fragment, may serve as a tool for gaining further insights into the domain structure and function of the myosin head.

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Section: Localization Of the Arg-c Clip Site On The S-1 Heavy Chainmentioning

confidence: 99%

Selective cleavage at lysine of the 50 kDa‐20 kDa connector loop segment of skeletal myosin S‐1 by endoproteinase Arg‐C

1989

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“…Chemical cross-linking of actin to SI resulted in a marked activation of Sl Mg2+-ATPase activity [6,7]. These results led to the notion that Sl and the smaller, 'outermost' N-terminal domain of actin [8,9] dock at multiple sites.…”

Section: Introductionmentioning

confidence: 99%

Peptide competition of actin activation of myosin‐subfragment 1 ATPase by an amino terminal actin fragment

et al. 1991

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The amino-terminal region of actin participates in the binding of myosin subfragment I (Sl) during cross-bridge cycling, thereby assisting in the activation of the magnesium-dependent myosin ATPase. Effects of three actin fragments on the magnesium-dependent Sl and acto-SI ATPase activities in solution were studied. One of the peptides, containing residues actin 1 44, mimicked the Sl ATPase-activating properties of actin and in turn inhibited acto-S I ATPasc both in a concentration-dependent manner. This suggests peptide competition for the actin binding site on myosin.The other f'ragments. residues actin 1 IX and X2 119, respectively, had no detectable effect on Sl-and acto-Sl ATPase activity.

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“…Thrombin treatment does not cleave M765 but degrades monomeric actin independently of the presence of myosin at residues 28, 39, and 113, leading to actin peptides 40-375 (A37), 114-375 (A27), 40-113 (A10), 1-28, and 29-39 (21,25,56,57). skeletal muscle myosin) with the actin-peptide 1-28 bound to the motor domain (21,58).…”

Section: Resultsmentioning

confidence: 99%

Functional Characterization of the Secondary Actin Binding Site of Myosin II

et al. 1999

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The role of the interaction between actin and the secondary actin binding site of myosin (segment 565-579 of rabbit skeletal muscle myosin, referred to as loop 3 in this work) has been studied with proteolytically generated smooth and skeletal muscle myosin subfragment 1 and recombinant Dictyostelium discoideum myosin II motor domain constructs. Carbodiimide-induced cross-linking between filamentous actin and myosin loop 3 took place only with the motor domain of skeletal muscle myosin and not with those of smooth muscle or D. discoideum myosin II. Chimeric constructs of the D. discoideum myosin motor domain containing loop 3 of either human skeletal muscle or nonmuscle myosin were generated. Significant actin cross-linking to the loop 3 region was obtained only with the skeletal muscle chimera both in the rigor and in the weak binding states, i.e., in the absence and in the presence of ATP analogues. Thrombin degradation of the cross-linked products was used to confirm the cross-linking site of myosin loop 3 within the actin segment 1-28. The skeletal muscle and nonmuscle myosin chimera showed a 4-6-fold increase in their actin dissociation constant, due to a significant increase in the rate for actin dissociation (k -A ) with no significant change in the rate for actin binding (k +A ). The actin-activated ATPase activity was not affected by the substitutions in the chimeric constructs. These results suggest that actin interaction with the secondary actin binding site of myosin is specific for the loop 3 sequence of striated muscle myosin isoforms but is apparently not essential either for the formation of a high affinity actinmyosin interface or for the modulation of actomyosin ATPase activity.Myosin molecules are actin-based molecular motors that use the energy supplied by the hydrolysis of ATP to perform various cell motility processes. The catalytic activity of myosin resides in its conserved motor domain, which interacts with actin, binds to and hydrolyzes ATP, and produces the force necessary for movement along actin filaments. Although the 3-D structure of the motor domain is highly conserved within the myosin family (1, 2), the enzymatic and motile activities of different myosin isoforms show a high degree of divergence (3). These functional variations are probably not due to fundamental differences in the molecular mechanism used by each isotype to convert the chemical energy into mechanical force but rather reflect a fine-tuning to specific functional requirements that is brought about by variations in the primary sequence of the motor domain itself. In agreement with this idea, sequence alignments reveal large differences in primary sequence at the proposed actin-myosin interface (4). Within this interface, one can clearly distinguish electrostatic and hydrophobic contacts (5, 6). The formation of the initial collision complex formed in the presence of ATP and ADP‚P i is largely an ionic interaction, whereas the transition from the weak to the strong binding complex has both ionic and hydro...

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Crosslinking of the skeletal myosin subfragment-1 heavy chain to the N-terminal actin segment of residues 40-113

Cited by 63 publications

References 52 publications

Selective cleavage at lysine of the 50 kDa‐20 kDa connector loop segment of skeletal myosin S‐1 by endoproteinase Arg‐C

Selective cleavage at lysine of the 50 kDa‐20 kDa connector loop segment of skeletal myosin S‐1 by endoproteinase Arg‐C

Peptide competition of actin activation of myosin‐subfragment 1 ATPase by an amino terminal actin fragment

Functional Characterization of the Secondary Actin Binding Site of Myosin II

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