Ono B, Naito K
Laboratory of Environmental Hygiene Chemistry, Faculty of Pharmaceutical Sciences, Okayama University, Japan.
Yeast. 1991 Nov;7(8):849-55. doi: 10.1002/yea.320070810.
Although Saccharomyces cerevisiae strains had different cysteine uptake activities, they revealed monophasic uptake kinetics and had the same KT (83.3 microM). The optimal pH of cysteine uptake was between 4.5 and 5.0, but the activity was quickly lost if cells were kept in buffer. When the activity was measured in the growth medium, it increased in the presence of EDTA and greatly decreased in the presence of mercuric chloride. Thioglycol as well as metabolic inhibitors such as dinitrophenol and azide were inhibitory. Homocysteine and methionine were competitive and non-competitive inhibitors, respectively. Cysteamine and cysteic acid were not inhibitory. From these observations, we conclude that the system mediating uptake of cysteine is specific (we thus name it the cysteine transport system) and that the cysteine transport system recognizes not only the SH-group but also amino- and carboxyl-groups. In wild-type strains the cysteine transport system was derepressed only when the cells were incubated without any sulfur source. On the other hand, in cysteine-dependent mutants, cysteine uptake activity increased with increase of exogenous supply of cysteine, glutathione or methionine. From this result, we suspect that the cellular cysteine level is the limiting factor for biosynthesis of the cysteine transport system in cysteine-dependent strains.
尽管酿酒酵母菌株具有不同的半胱氨酸摄取活性,但它们呈现出单相摄取动力学,且具有相同的转运常数(83.3微摩尔)。半胱氨酸摄取的最适pH在4.5至5.0之间,但如果将细胞保存在缓冲液中,活性会迅速丧失。当在生长培养基中测量活性时,在乙二胺四乙酸(EDTA)存在下活性增加,而在氯化汞存在下活性大大降低。硫代乙醇以及代谢抑制剂如二硝基苯酚和叠氮化物具有抑制作用。同型半胱氨酸和甲硫氨酸分别是竞争性和非竞争性抑制剂。半胱胺和半胱氨酸无抑制作用。从这些观察结果中,我们得出结论,介导半胱氨酸摄取的系统具有特异性(因此我们将其命名为半胱氨酸转运系统),并且半胱氨酸转运系统不仅识别巯基,还识别氨基和羧基。在野生型菌株中,只有当细胞在没有任何硫源的情况下孵育时,半胱氨酸转运系统才会去阻遏。另一方面,在半胱氨酸依赖型突变体中,半胱氨酸摄取活性随着半胱氨酸、谷胱甘肽或甲硫氨酸外源供应的增加而增加。根据这一结果,我们推测细胞内半胱氨酸水平是半胱氨酸依赖型菌株中半胱氨酸转运系统生物合成的限制因素。