Butler A R, O'Donnell R W, Martin V J, Gooday G W, Stark M J
Department of Biochemistry, The University, Dundee, Scotland.
Eur J Biochem. 1991 Jul 15;199(2):483-8. doi: 10.1111/j.1432-1033.1991.tb16147.x.
The Kluyveromyces lactis toxin is a protein containing three subunits (alpha, beta and gamma) which causes sensitive yeast cells to arrest proliferation in the G1 phase of the cell cycle. Despite the toxin's complex structure, the gamma subunit appears to be the only component required for it to arrest proliferation since intracellular expression of the gamma polypeptide alone in a sensitive yeast strain mimics the effect of the exogenous native toxin. The toxin alpha subunit shows sequence similarity to a variety of chitinases and here we report that the toxin is a potent exochitinase. The exochitinase activity is absolutely required for its biological activity against sensitive Saccharomyces cerevisiae cells and allosamidin, a specific inhibitor of chitinases, abolishes the biological activity of the toxin. However, since the alpha subunit is not required for the G1 arrest induced by the toxin, the chitinase activity of the toxin cannot be directly responsible for the ultimate effect of the toxin and most likely plays a role in the initial interaction of the toxin with sensitive cells.
乳酸克鲁维酵母毒素是一种包含三个亚基(α、β和γ)的蛋白质,它会导致敏感酵母细胞在细胞周期的G1期停止增殖。尽管毒素结构复杂,但γ亚基似乎是其阻止增殖所需的唯一成分,因为仅在敏感酵母菌株中细胞内表达γ多肽就能模拟外源性天然毒素的作用。毒素α亚基与多种几丁质酶具有序列相似性,我们在此报告该毒素是一种强效外切几丁质酶。其外切几丁质酶活性对于其针对敏感酿酒酵母细胞的生物活性是绝对必需的,而几丁质酶的特异性抑制剂别洛沙米定可消除该毒素的生物活性。然而,由于毒素诱导的G1期停滞并不需要α亚基,所以毒素的几丁质酶活性不能直接导致毒素的最终作用,很可能在毒素与敏感细胞的初始相互作用中发挥作用。