Blair D F, Berg H C
Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
Cell. 1990 Feb 9;60(3):439-49. doi: 10.1016/0092-8674(90)90595-6.
A number of mutants of motA, a gene necessary for flagellar rotation in E. coli, were isolated and characterized. Many mutations were dominant, owing to competition between functional and nonfunctional MotA for a limited number of sites on the flagellar motor. A new class of mutant was discovered in which flagellar torque is normal at low speeds but reduced at high speeds. Hydrogen isotope effects on these mutants indicate that MotA catalyzes proton transfer. We confirmed an earlier observation that overproduction of MotA leads to accumulation of the protein in the cytoplasmic membrane and to significant decreases in growth rate. When nonfunctional mutant variants of MotA were overproduced instead, they accumulated in the cytoplasmic membrane, but growth was not impaired. These results also suggest that MotA conducts protons. This was confirmed by measuring the proton permeabilities of vesicles containing wild-type or mutant MotA proteins.
分离并鉴定了大肠杆菌鞭毛旋转所必需的motA基因的多个突变体。许多突变是显性的,这是由于功能性和非功能性MotA在鞭毛马达上有限数量的位点上存在竞争。发现了一类新的突变体,其鞭毛扭矩在低速时正常,但在高速时降低。氢同位素对这些突变体的影响表明MotA催化质子转移。我们证实了早期的一项观察结果,即MotA的过量表达会导致该蛋白在细胞质膜中积累,并导致生长速率显著下降。相反,当MotA的无功能突变变体过量表达时,它们会在细胞质膜中积累,但生长并未受到损害。这些结果还表明MotA传导质子。通过测量含有野生型或突变型MotA蛋白的囊泡的质子渗透性,证实了这一点。