Li Yuezhou, Moe Paul C, Chandrasekaran Subramanian, Booth Ian R, Blount Paul
Department of Physiology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9040, USA.
EMBO J. 2002 Oct 15;21(20):5323-30. doi: 10.1093/emboj/cdf537.
Three gene products that form independent mechanosensitive channel activities have been identified in Escherichia coli. Two of these, MscL and MscS, play a vital role in allowing the cell to survive acute hypotonic stress. Much less is known of the third protein, MscK (KefA). Here, we characterize the MscK channel activity and compare it with the activity of its structural and functional homologue, MscS. While both show a slight anionic preference, MscK appears to be more sensitive to membrane tension. In addition, MscK, but not MscS activity appears to be regulated by external ionic environment, requiring not only membrane tension but also high concentrations of external K(+), NH(4)(+), Rb(+) or Cs(+) to gate; no activity is observed with Na(+), Li(+) or N-methyl-D-glucamine (NMDG). An MscK gain-of-function mutant gates spontaneously in the presence of K(+) or similar ions, and will gate in the presence of Na(+), Li(+) and NMDG, but only when stimulated by membrane tension. Increased sensitivity and the highly regulated nature of MscK suggest a more specialized physiological role than other bacterial mechanosensitive channels.
在大肠杆菌中已鉴定出三种形成独立机械敏感通道活性的基因产物。其中两种,即MscL和MscS,在使细胞在急性低渗应激下存活方面发挥着至关重要的作用。对于第三种蛋白质MscK(KefA)的了解则少得多。在此,我们对MscK通道活性进行了表征,并将其与其结构和功能同源物MscS的活性进行了比较。虽然两者都表现出对阴离子的轻微偏好,但MscK似乎对膜张力更敏感。此外,MscK的活性似乎受外部离子环境的调节,而MscS则不然,MscK不仅需要膜张力,还需要高浓度的外部K⁺、NH₄⁺、Rb⁺或Cs⁺来开启通道;在Na⁺、Li⁺或N - 甲基 - D - 葡糖胺(NMDG)存在时未观察到活性。一个MscK功能获得性突变体在K⁺或类似离子存在时会自发开启通道,并且在Na⁺、Li⁺和NMDG存在时也会开启通道,但仅在受到膜张力刺激时才会开启。MscK更高的敏感性和高度受调节的特性表明其生理作用比其他细菌机械敏感通道更为特殊。