The Kinetic Mechanism of Myo1e (Human Myosin-IC)*

The Kinetic Mechanism of Myo1e (Human Myosin-IC)*
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Myo1e(人肌球蛋白-IC)的动力学机制*

DOI:
--
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发表时间:
2002
影响因子:
4.8
通讯作者:
E. Ostap
E. Ostap
中科院分区:
生物学2区
文献类型:
--
作者:
M. Mezgueldi;N. Tang;S. Rosenfeld;E. Ostap

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Myo 1 e是广泛表达的肌球蛋白-I家族的亚类-1成员。我们进行了动力学分析的截断myo 1 e,由电机和单一的智商基序与绑定钙调素。我们确定了ATP酶循环中关键步骤的速率和平衡常数。肌动蛋白激活ATP酶的最大速率(Vmax)和半最大速率时的肌动蛋白浓度(K-ATP酶)与天然蛋白相似。ATP酶的K值很低(约1 μm),但在ATP存在下,肌动蛋白与肌酶的亲和力很弱。弱的肌动蛋白亲和力和快速的磷酸盐释放速率导致在体外测定条件下磷酸盐被释放而myo 1 e从肌动蛋白解离的途径。肌动蛋白激活ATP酶活性和低K ATP酶是肌动蛋白激活ADP释放的结果。我们提出,myo 1 e被调谐到高浓度的交联肌动蛋白丝的区域中发挥作用。此外,我们发现肌动蛋白释放ADP的速度比其他脊椎动物肌球蛋白I亚型快10倍以上。我们建议,亚类-1肌球蛋白是调整快速滑动,而亚类-2亚型调整张力维持或应力传感。
Myo1e is the widely expressed subclass-1 member of the myosin-I family. We performed a kinetic analysis of a truncated myo1e that consists of the motor and the single IQ motif with a bound calmodulin. We determined the rates and equilibrium constants for the key steps in the ATPase cycle. The maximum actin activated ATPase rate (V max) and the actin concentration at half-maximum of V max(K ATPase) of myo1e are similar to those of the native protein. The K ATPase is low (∼1 μm), however the affinity of myo1e for actin in the presence of ATP is very weak. A weak actin affinity and a rapid rate of phosphate release result in a pathway under in vitro assay conditions in which phosphate is released while myo1e is dissociated from actin. Actin activation of the ATPase activity and the lowK ATPase are the result of actin activation of ADP release. We propose that myo1e is tuned to function in regions of high concentrations of cross-linked actin filaments. Additionally, we found that ADP release from actomyo1e is > 10-fold faster than other vertebrate myosin-I isoforms. We propose that subclass-1 myosin-Is are tuned for rapid sliding, whereas subclass-2 isoforms are tuned for tension maintenance or stress sensing.
使用新型磷酸盐荧光探针测量相关兔骨骼肌动球蛋白的核苷三磷酸裂解和磷酸盐释放步骤的动力学。
DOI: 10.1021/bi970540h
发表时间: 1997
期刊: Biochemistry.
影响因子: --
作者:
White,HD;Belknap,B;Webb,MR
通讯作者: Webb,MR
DOI: 10.1073/pnas.96.24.13726
发表时间: 1999-11-23
影响因子: 11.1
作者:
De La Cruz, EM;Wells, AL;Sweeney, HL
通讯作者: Sweeney, HL
DOI: 10.1091/mbc.12.4.780
发表时间: 2001-04-01
影响因子: 3.3
作者:
Berg, JS;Powell, BC;Cheney, RE
通讯作者: Cheney, RE
DOI: 10.1073/pnas.82.3.658
发表时间: 1985-01-01
影响因子: 11.1
作者:
SIEMANKOWSKI, RF;WISEMAN, MO;WHITE, HD
通讯作者: WHITE, HD
源自鸡钙调蛋白 cDNA 和钙调蛋白加工基因的蛋白质的细菌表达和表征。
DOI: --
发表时间: 1985
期刊: The Journal of biological chemistry
影响因子: --
作者:
Putkey,JA;Slaughter,GR;Means,AR
通讯作者: Means,AR