Department of Biophysics, University of Osnabrück, 49069 Osnabrück, Germany.
Proc Natl Acad Sci U S A. 2011 Mar 8;108(10):3924-9. doi: 10.1073/pnas.1011581108. Epub 2011 Feb 22.
ATP is synthesized by ATP synthase (F(O)F(1)-ATPase). Its rotary electromotor (F(O)) translocates protons (in some organisms sodium cations) and generates torque to drive the rotary chemical generator (F(1)). Elastic power transmission between F(O) and F(1) is essential for smoothing the cooperation of these stepping motors, thereby increasing their kinetic efficiency. A particularly compliant elastic domain is located on the central rotor (c(10-15)/ε/γ), right between the two sites of torque generation and consumption. The hinge on the active lever on subunit β adds further compliance. It is under contention whether or not the peripheral stalk (and the "stator" as a whole) also serves as elastic buffer. In the enzyme from Escherichia coli, the most extended component of the stalk is the homodimer b(2), a right-handed α-helical coiled coil. By fluctuation analysis we determined the spring constant of the stator in response to twisting and bending, and compared wild-type with b-mutant enzymes. In both deformation modes, the stator was very stiff in the wild type. It was more compliant if b was elongated by 11 amino acid residues. Substitution of three consecutive residues in b by glycine, expected to destabilize its α-helical structure, further reduced the stiffness against bending deformation. In any case, the stator was at least 10-fold stiffer than the rotor, and the enzyme retained its proton-coupled activity.
ATP 由 ATP 合酶(F(O)F(1)-ATP 酶)合成。其旋转电动(F(O)) 转运质子(在一些生物体中是钠离子)并产生扭矩来驱动旋转化学发生器(F(1))。F(O) 和 F(1) 之间的弹性动力传递对于平滑这些步进电机的协作至关重要,从而提高它们的动力效率。一个特别有弹性的弹性域位于中央转子(c(10-15)/ε/γ)上,正好在两个扭矩产生和消耗的位置之间。亚基β上的活性杆的铰链增加了进一步的弹性。外周柄(以及作为整体的“定子”)是否也作为弹性缓冲器存在争议。在大肠杆菌中的酶中,柄的最延伸部分是同源二聚体 b(2),一种右手α-螺旋卷曲螺旋。通过波动分析,我们确定了定子在扭转和弯曲时的弹性常数,并将野生型与 b-突变酶进行了比较。在这两种变形模式中,定子在野生型中非常坚硬。如果 b 延长 11 个氨基酸残基,则会增加其弹性。用甘氨酸替代 b 中的三个连续残基,预计会破坏其α-螺旋结构,进一步降低弯曲变形的刚度。在任何情况下,定子的刚度至少比转子高 10 倍,并且酶保留其质子偶联活性。