scholarly journals Protein fluorescence changes associated with ATP and adenosine 5′-[γ-thio]triphosphate binding to skeletal muscle myosin subfragment 1 and actomyosin subfragment 1

1988 ◽  
Vol 249 (3) ◽  
pp. 735-743 ◽  
Author(s):  
N C Millar ◽  
M A Geeves
Keyword(s):  
Indirect Evidence ◽  
Protein Fluorescence ◽  
Skeletal Muscle Myosin ◽  
Myosin Subfragment ◽  
Fluorescence Change ◽  
Myosin Subfragment 1 ◽  
Cleavage Step ◽  
Muscle Myosin ◽  
Gamma S ◽  
Dissociated State

1. The fluorescence changes accompanying the binding of ATP and adenosine 5′-[gamma-thio]triphosphate (ATP gamma S) to myosin subfragment 1 (S1) and actomyosin subfragment 1 (actoS1) have been reinvestigated at 20 degrees C and 1 degree C, pH 7.0, 0.1 M-KCl. 2. Two successive fluorescence enhancements are observed following ATP binding to both S1 and actoS1. 3. The slow fluorescence change has the same rate with S1 and actoS1, and is due to the ATP cleavage step. 4. With actoS1 the fast fluorescence change occurs after dissociation, so a new intermediate, S1 ATP, is required on the actoS1 pathway. 5. The dissociation of actoS1 by ATP gamma S results in a fluorescence enhancement with the same apparent rate as dissociation, but indirect evidence suggests that this too occurs on a dissociated state.

scholarly journals Correlation between calponin and myosin subfragment 1 binding to F-actin and ATPase inhibition

1997 ◽  
Vol 321 (2) ◽  
pp. 519-523 ◽  
Author(s):  
Pawel T. SZYMANSKI ◽  
Zenon GRABAREK ◽  
Terence TAO
Keyword(s):  
Troponin I ◽  
Sequence Similarity ◽  
Actin Binding ◽  
Amino Acid Sequence Similarity ◽  
Smooth Muscle Contractility ◽  
Skeletal Muscle Myosin ◽  
Myosin Subfragment ◽  
Myosin Subfragment 1 ◽  
Muscle Myosin ◽  
Atpase Inhibition

Calponin is a thin-filament-associated protein that has been implicated in the regulation of smooth-muscle contractility. It binds to F-actin and inhibits the MgATPase activity of actomyosin. In the present work we have examined the effect of recombinant chicken gizzard α-calponin (RαCaP) on the binding of rabbit skeletal-muscle myosin subfragment 1 (S1) to F-actin and on the inhibition of its actin-activated MgATPase. We have found that binding of one RαCaP molecule to every three to four actin monomers is sufficient for maximal inhibition of actoŐS1 ATPase. At this RαCaP/actin ratio RαCaP does not interfere with S1 binding to F-actin. At higher concentrations, RαCaP displaces S1 from F-actin and a 1:1 RαCaPŐactin monomer complex is formed. RαCaP is also able to displace troponin I from its complex with F-actin which may reflect the amino acid sequence similarity between RαCaP and troponin I in their actin-binding regions.

Skeletal muscle myosin subfragment 1 dimers

1997 ◽  
Vol 65 (1) ◽  
pp. 85-90 ◽  
Author(s):  
Karen Claire ◽  
Robert Pecora ◽  
Stefan Highsmith
Keyword(s):  
Skeletal Muscle ◽  
Skeletal Muscle Myosin ◽  
Myosin Subfragment ◽  
Myosin Subfragment 1 ◽  
Muscle Myosin

scholarly journals Characterization of an actin-myosin head interface in the 40–113 region of actin using specific antibodies as probes

1990 ◽  
Vol 271 (2) ◽  
pp. 407-413 ◽  
Author(s):  
J P Labbé ◽  
C Méjean ◽  
Y Benyamin ◽  
C Roustan
Keyword(s):  
Myosin Head ◽  
Interface Model ◽  
Filamentous Actin ◽  
Skeletal Muscle Myosin ◽  
Myosin Subfragment ◽  
Myosin Subfragment 1 ◽  
Myosin Interaction ◽  
Muscle Myosin ◽  
Covalent Complex

Evidence for the participation of the 1-7 and 18-28 N-terminal sequences of actin at different steps of actin-myosin interaction process is well documented in the literature. Cross-linking of the rigor complex between filamentous actin and skeletal-muscle myosin subfragment 1 was accomplished by the carboxy-group-directed zero-length protein cross-linker, 1-ethyl-3-[3-(dimethylamino)propyl]carbodi-imide. After chaotropic depolymerization and thrombin digestion, which cleaves only actin, the covalent complex with Mr 100,000 was characterized by PAGE. The linkage was identified as being between myosin subfragment 1 (S-1) heavy chain and actin-(1-28)-peptide. The purified complex retained in toto its ability to combine reversibly with fresh filamentous actin, but showed a decrease in the Vmax. of actin-dependent Mg2(+)-ATPase. By using e.l.i.s.a., S-1 was observed to bind to coated monomeric actin or its 1-226 N-terminal peptide. This interaction strongly interfered with the binding of antibodies directed against the 95-113 actin sequence. Moreover, S-1 was able to bind with coated purified actin-(40-113)-peptide. Finally, antibodies directed against the 18-28 and 95-113 actin sequence, which strongly interfered with S1 binding, were unable to compete with each other. These results suggest that two topologically independent regions are involved in the actin-myosin interface: one located in the conserved 18-28 sequence and the other near residues 95-113, including the variable residue at position 89. Other experiments support the ‘multisite interface model’, where the two actin sites could modulate each other during S-1 interaction.

scholarly journals Interaction of skeletal-muscle myosin subfragment-1 with the actin-(338-348) peptide.

1994 ◽  
Vol 299 (3) ◽  
pp. 875-879 ◽  
Author(s):  
J P Labbé ◽  
S Lelievre ◽  
M Boyer ◽  
Y Benyamin
Keyword(s):  
Skeletal Muscle ◽  
Binding Site ◽  
Antigenic Site ◽  
Filamentous Actin ◽  
Apparent Dissociation Constant ◽  
Skeletal Muscle Myosin ◽  
Myosin Subfragment ◽  
Myosin Subfragment 1 ◽  
Order Of Magnitude ◽  
Muscle Myosin

The data presented here confirm and provide further experimental evidence that rabbit skeletal-muscle myosin subfragment-1 (S-1) binds to the postulated actin-(338-348) hydrophobic segment [Kabsch, Mannherz, Suck, Pai and Holmes (1990) Nature (London) 347, 37-44] with high affinity in the absence and presence of MgATP. The apparent dissociation constant of the S-1 interaction (5.5 x 10(-7) M) with the actin-(338-348) peptide was of the same order of magnitude as that of the actin-(18-28) binding site (2 x 10(-6) M). In similar conditions, fragmented (27 kDa-50 kDa-20 kDa) S-1 also bound to the peptide. Antibodies directed to the vicinal sequence 348-358 were rapidly eliminated from actin by S-1 interaction and weakened S-1 binding to monomeric or filamentous actin. The antigenic site (348-358) is located very close to the C-terminal S-1-binding site (360-369) and encompasses some residues (Leu-349 and Phe-352) included in the hydrophobic S-1-binding region [Schröder, Manstein, Jahn, Holden, Rayment, Holmes and Spudich (1993) Nature (London) 364, 171-174]. It was observed that anti-[actin-(348-358)] antibodies were also unable to decrease actomyosin ATPase activity, in contrast with previous results obtained with anti-[actin-(18-28)] antibodies [Adams and Reisler (1993) Biochemistry 32, 5051-5056]. The hydrophobic actin-(338-348) peptide used in considerable excess was unable to perturb acto-S-1 and S-1 activities in contrast with results obtained with the N-terminal actin peptide [Kôgler, Moir, Trayer and Ruegg (1991) FEBS Lett. 294, 31-34].

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