Measurement of the fraction of myosin heads bound to actin in rabbit skeletal myofibrils in rigor.

Measurement of the fraction of myosin heads bound to actin in rabbit skeletal myofibrils in rigor.
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测量兔骨骼肌原纤维中与肌动蛋白结合的肌球蛋白头部的分数。

DOI:
10.1016/0022-2836(81)90352-1
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发表时间:
1981
影响因子:
5.6
通讯作者:
Harrington,WF
Harrington,WF
中科院分区:
生物学2区
文献类型:
--
作者:
Lovell,SJ;Harrington,WF

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在生理离子强度和pH下,兔骨骼肌原纤维的胰蛋白酶消化导致肌球蛋白重链在十二烷基硫酸钠/聚丙烯酰胺凝胶上的一个位点裂解,产生两条带(Mr= 200,000和26,000)。加入焦磷酸钠(至1 mm)使肌球蛋白头从肌动蛋白上解离后,胰蛋白酶蛋白水解导致产生三条带,160 K、51 K和26 K,其中74 K带作为51 K和26 K物质的前体。在这些条件下,重链向重和轻裂肌球蛋白的裂解不显著。当用木瓜蛋白酶消化时,胰蛋白酶消化的肌原纤维产生与天然肌原纤维相同量的杆。这些结果表明,肌动蛋白阻断胰蛋白酶切割的肌球蛋白重链在一个网站74 K从N端。从51 K物质的消化形成的僵硬纤维在不同的肌节长度的量的测量,我们估计,至少95%的肌球蛋白头绑定到肌动蛋白在100%重叠的厚和薄的细丝。因此,所有的肌球蛋白分子都可以与肌动蛋白结合,因此在严格的条件下,肌球蛋白分子的两个头可以同时与肌动蛋白丝相互作用。
Tryptic digestion of rabbit skeletal myofibrils at physiological ionic strength and pH results in cleavage of the myosin heavy chain at one site giving two bands (Mr= 200,000 and 26,000) on sodium dodecyl sulfate/polyacrylamide gels. Following addition of sodium pyrophosphate (to 1 mm) to dissociate the myosin heads from actin, tryptic proteolysis results in production of three bands, 160K†, 51K and 26K, with a 74K band appearing as a precursor of the 51K and 26K species. Under these conditions, there is insignificant cleavage of heavy chain to the heavy and light meromyosins. Trypsin-digested myofibrils yield the same amount of rod as native myofibrils when digested with papain. These results indicate that actin blocks tryptic cleavage of the myosin heavy chain at a site 74K from the N terminus. From measurements of the amount of 51K species formed by digestion of rigor fibers at various sarcomere lengths, we estimate that at least 95% of the myosin heads are bound to actin at 100% overlap of thick and thin filaments. Hence all myosin molecules can bind to actin, and consequently both heads of a myosin molecule can interact simultaneously with actin filaments under rigor conditions.
DOI: 10.1016/s0022-2836(67)80046-9
发表时间: 1967-01-01
影响因子: 5.6
作者:
HUXLEY, HE;BROWN, W
通讯作者: BROWN, W
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DOI: --
发表时间: 1974
影响因子: 3.9
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DOI: --
发表时间: 1978
影响因子: 5.6
作者:
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通讯作者: W. F. Harrington
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发表时间: 1964
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影响因子: --
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