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Dimerization of native myosin LC2(RLC)-free subfragment 1 from adult rabbit skeletal muscle

机译:成年兔骨骼肌中不含天然肌球蛋白LC2(RLC)的亚片段1的二聚化

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We reinvestigated whether the native myosin LC2-free-subfragment 1 (S1) dimer exists by using viscometry, capillary electrophoresis, and laser light scattering. We found that the intrinsic viscosity of the monomer is [eta](m) = 6.7 cm(3)/g and its translation diffusion coefficient is D-t,m(20,w)(c = 0) = 4.43 x 10(-7) cm(2)/s. For the dimer, [eta](d) = 19.8 cm(3)/g and D-t,d(20,w)(c = 0) = 2.54 x 10(-7) cm(2)/s. Using the Svedberg equation and introducing the values of the sedimentation coefficients (5.05 S for the monomer and 6.05 S for the dimer), we find the following molecular weights: M-r,M-m = 108 000 Da and M-r,M-d = 213 000 Da, which agree well with previous determinations. Capillary electrophoresis successfully separated S1(A1) and S1(A2), in a monomer buffer, and SI(AI) and S1(A2) and a heterodimer S1(AI)S1(A2), in a dimer buffer. An interesting feature of the monomer-dimer equilibrium is the presence of temperature transitions, whose positions and widths depend upon the buffer conditions. At low temperatures, a pure dimer was observed, whereas at high temperatures only the monomer was present. The dimerization site on both myosin and S1 is extremely labile. [References: 29]
机译:我们使用粘度测定法,毛细管电泳和激光散射技术重新研究了天然肌球蛋白LC2游离亚片段1(S1)二聚体是否存在。我们发现单体的特性粘度为η(m)= 6.7cm(3)/ g,并且其平移扩散系数为Dt,m(20,w)(c = 0)= 4.43×10(-7)。 )cm(2)/秒。对于二聚体,η(d)= 19.8cm(3)/ g,D-t,d(20,w)(c = 0)= 2.54×10(-7)cm(2)/ s。使用Svedberg方程并引入沉降系数的值(单体为5.05 S,二聚体为6.05 S),我们发现以下分子量:Mr,Mm = 108000 Da和Mr,Md = 213000 Da,其中与先前的决定非常吻合。毛细管电泳在单体缓冲液中成功分离了S1(A1)和S1(A2),在二聚体缓冲液中成功分离了SI(AI)和S1(A2)和异二聚体S1(AI)S1(A2)。单体-二聚体平衡的一个有趣特征是存在温度跃迁,其位置和宽度取决于缓冲条件。在低温下,观察到纯二聚体,而在高温下仅存在单体。肌球蛋白和S1上的二聚位点非常不稳定。 [参考:29]

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