Uenoyama Atsuko, Miyata Makoto
Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585, Japan.
Proc Natl Acad Sci U S A. 2005 Sep 6;102(36):12754-8. doi: 10.1073/pnas.0506114102. Epub 2005 Aug 26.
Several species of mycoplasmas glide on solid surfaces, in the direction of their membrane protrusion at a cell pole, by an unknown mechanism. Our recent studies on the fastest species, Mycoplasma mobile, suggested that the gliding machinery, localized at the base of the membrane protrusion (the "neck"), is composed of two huge proteins. This machinery forms spikes sticking out from the neck and propels the cell by alternately binding and unbinding the spikes to a solid surface. Here, to study the intracellular mechanisms for gliding, we established a permeabilized gliding ghost model, analogous to the "Triton model" of the eukaryotic axoneme. Treatment with Triton X-100 stopped the gliding and converted the cells to permeabilized "ghosts." When ATP was added exogenously, approximately 85% of the ghosts were reactivated, gliding at speeds similar to those of living cells. The reactivation activity and inhibition by various nucleotides and ATP analogs, as well as their kinetic parameters, showed that the machinery is driven by the hydrolysis of ATP to ADP plus phosphate, caused by an unknown ATPase.
几种支原体能够在固体表面沿着细胞膜在细胞极处的突出方向滑行,但其机制尚不清楚。我们最近对滑行速度最快的物种——运动支原体的研究表明,位于细胞膜突出部基部(“颈部”)的滑行机制由两种巨大的蛋白质组成。这种机制形成从颈部伸出的尖刺,并通过使尖刺与固体表面交替结合和解离来推动细胞。在此,为了研究滑行的细胞内机制,我们建立了一种通透化的滑行鬼模型,类似于真核生物轴丝的“Triton模型”。用Triton X-100处理会使滑行停止,并将细胞转化为通透化的“鬼细胞”。当外源添加ATP时,约85%的鬼细胞被重新激活,以与活细胞相似的速度滑行。各种核苷酸和ATP类似物的重新激活活性和抑制作用以及它们的动力学参数表明,该机制由一种未知的ATP酶催化ATP水解为ADP和磷酸所驱动。