Ding P Z, Botfield M C, Wilson T H
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Biochim Biophys Acta. 2000 Dec 20;1509(1-2):123-30. doi: 10.1016/s0005-2736(00)00286-8.
Melibiose carrier mutants, isolated by growing cells on melibiose plus the non-metabolizable competitive inhibitor thiomethyl-beta-galactoside (TMG), were studied to determine sugar and cation recognition abnormalities. Most of the mutants show good transport of melibiose but have lost the recognition of TMG. In addition, most mutants show little or no transport of lactose. Cation recognition is also affected as all of these mutants have lost the ability to transport protons with melibiose. The amino acids causing these mutations were determined by sequencing the melB gene on the plasmid. The mutations were located on helices I, IV, VII, X and XI. We propose that these five helices are in proximity with each other and that they line the sugar/cation transport channel.
通过在蜜二糖加上不可代谢的竞争性抑制剂硫代甲基-β-半乳糖苷(TMG)上培养细胞分离得到的蜜二糖载体突变体,被用于研究糖和阳离子识别异常情况。大多数突变体表现出良好的蜜二糖转运能力,但失去了对TMG的识别能力。此外,大多数突变体对乳糖的转运很少或完全没有。阳离子识别也受到影响,因为所有这些突变体都失去了与蜜二糖一起转运质子的能力。通过对质粒上的melB基因进行测序确定了导致这些突变的氨基酸。这些突变位于螺旋I、IV、VII、X和XI上。我们提出这五个螺旋彼此靠近,并且它们构成了糖/阳离子转运通道的内壁。