The proteolytic substructure of light meromyosin. Localization of a region responsible for the low ionic strength insolubility of myosin.

Nyitray, László; Mócz, Gábor; Szilágyi, László; Bálint, M.; Lu, Renne Chen; Wong, Anna; Gergely, J.

doi:10.1016/s0021-9258(17)44103-2

Cited by 66 publications

(13 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has been claimed that LMM fragments with different molecular weights from carp fast skeletal muscle have differences in the C-terminal length(17). Additional differences probably result from multiple proteolytic cleavage sites in the LMM: subfragment-2 junction of myosin rod as in the case of rabbit fast skeletal muscle(25). In relation to these facts, we have observed that myosin rods prepared from the 10 and 30 °Cacclimated carp show no difference in R-chymotryptic susceptibility(26).…”

mentioning

confidence: 47%

Differential Scanning Calorimetry and CD Spectrometry of Acclimation Temperature-Associated Types of Carp Light Meromyosin

et al. 1997

View full text Add to dashboard Cite

Differential scanning calorimetry and CD spectrometry were employed to study the thermal unfolding of light meromyosin (LMM) prepared from carp acclimated to different temperatures. The transition temperatures given by the major peaks at pH 8.0 in 0.6 M KCl for LMM from carp acclimated to 10 degrees C were 32.5 and 39.5 degrees C with the calorimetric enthalpies (DeltaHcal) of 269 and 52 kcal/mol, respectively. LMM from carp acclimated to 20 degrees C exhibited three peaks of transition temperatures at 34.5, 40.2, and 46.9 with DeltaHcal of 152, 20, and 10 kcal/mol, respectively. On the other hand, LMM from carp acclimated to 30 degrees C showed two different patterns. The first experiment gave two transition temperatures at 39.2 and 47.3 degrees C with DeltaHcal of 231 and 39 kcal/mol, respectively. The second series of experiments resulted in showing three peaks of 34.4, 39.5, and 47.5 degrees C with DeltaHcal of 117, 123, and 28 kcal/mol, respectively. N-terminal amino acid sequence analysis revealed that LMM at the second series of experiments with the 30 degrees C-acclimated carp contained component(s) predominant in the 20 degrees C-acclimated carp. Thermal unfolding responsible for these transition temperatures was well explained by melting of alpha-helices which could be determined by far-ultraviolet CD spectroscopy. These results clearly demonstrate that the 30 degrees C-acclimated carp contained the most thermostable LMM.

show abstract

mentioning

confidence: 47%

Differential Scanning Calorimetry and CD Spectrometry of Acclimation Temperature-Associated Types of Carp Light Meromyosin

et al. 1997

View full text Add to dashboard Cite

show abstract

“…The effect of these antibodies on polymerization is consistent with the observation that the COOHterminal region of the rod is important for filament assembly in a number of myosins. The aggregation of skeletal LMM at low ionic strength is due to a small region located '~15-20 nm from the COOH terminus of the skeletal muscle rod (Nyitray et al, 1983). A larger, 20-nm COOH-terminal segment, which overlaps the region responsible for insolubility in skeletal myosin, appears to play a role in intermolecular interactions between smooth muscle myosin rods (Cross and Vandekerckhove, 1986).…”

Section: Discussionmentioning

confidence: 98%

Monoclonal antibodies detect and stabilize conformational states of smooth muscle myosin.

Trybus

Henry

1989

The Journal of Cell Biology

View full text Add to dashboard Cite

Abstract. Antibodies with epitopes near the heavy meromyosin/light meromyosin junction distinguish the folded from the extended conformational states of smooth muscle myosin. Antibody 10S.1 has 100-fold higher avidity for folded than for extended myosin, while antibody S2.2 binds preferentially to the extended state. The properties of these antibodies provide direct evidence that the conformation of the rod is different in the folded than the extended monomeric state, and suggest that this perturbation may extend into the subfragment 2 region of the rod. Two antihead antibodies with epitopes on the heavy chain map at or near the head/rod junction. Magnesium greatly enhances the binding of these antibodies to myosin, showing that the conformation of the heavy chain in the neck region changes upon divalent cation binding to the regulatory light chain. Myosin assembly is also altered by antibody binding. Antibodies that bind to the central region of the rod block disassembly of iliamerits upon MgATP addition. Antibodies with epitopes near the COOH terminus of the rod, in contrast, promote filament depolymerization, suggesting that this region of the tail is important for assembly. The monoctonal antibodies described here are therefore useful both for detecting and altering conformational states of smooth muscle myosin.

show abstract

“…Fór LMM the answer is very easy: In conventionally prepared proteolytic fragments of myosin, it is extremely likely that the mobile C-terminus is cleaved off during the digestion [see, e.g., Nyitray et al (1983)]. For myosin itself the answer is probably that nobody did look for such sharp lines, that one needs a very good signal-to-noise ratio with rather low concentrations of myosin (low viscosity of the solution!…”

Section: Discussionmentioning

confidence: 99%

C-terminal structure and mobility of rabbit skeletal muscle light meromyosin as studied by one- and two-dimensional proton NMR spectroscopy and x-ray small-angle scattering

Kalbitzer¹,

Maéda²,

Rösch³

et al. 1991

Biochemistry

View full text Add to dashboard Cite

Intact rabbit myosin and two different C-terminal fragments of rabbit muscle light meromyosin (LMM) expressed in Escherichia coli, LMM-30, and LMM-30C', were studied by 1H NMR spectroscopy. X-ray small-angle scattering shows that at high ionic strength two polypeptide chains of LMM-30 (which consists of the C-terminal 262 amino acids of myosin heavy chain) or LMM-30C' (which corresponds to LMM-30 but lacks the last 17 residues) assemble to form an alpha-helical coiled-coil as it is found also in myosin. The last 12 C-terminal residues of one polypeptide chain of LMM-30 and the last 9 C-terminal residues of the other chain are very mobile. The last 8 residues of the two strands are equivalent from the NMR point of view and unfolded; the valine residues in position 255 in the two strands are not equivalent, suggesting an interaction between the two strands, Ser-252, Arg-253, and Asp-254 are completely immobilized in one of the polypeptide strands and partly mobile in the other. Essentially the same pattern is observed in intact myosin. In spite of the large molecular weights of LMM-30 and LMM-30C', it is possible to resolve almost all aromatic residues and to determine the pK values of all the 4 tyrosine and of 9 (out of 10) histidine residues. The tyrosine residues in the two strands are equivalent in the two polypeptide chains and both have a pK of 10.5. The pK values of the histidine residues vary between 5.7 and 7.0.

show abstract

The proteolytic substructure of light meromyosin. Localization of a region responsible for the low ionic strength insolubility of myosin.

Cited by 66 publications

References 34 publications

Differential Scanning Calorimetry and CD Spectrometry of Acclimation Temperature-Associated Types of Carp Light Meromyosin

Differential Scanning Calorimetry and CD Spectrometry of Acclimation Temperature-Associated Types of Carp Light Meromyosin

Monoclonal antibodies detect and stabilize conformational states of smooth muscle myosin.

C-terminal structure and mobility of rabbit skeletal muscle light meromyosin as studied by one- and two-dimensional proton NMR spectroscopy and x-ray small-angle scattering

Contact Info

Product

Resources

About