Vandenberghe Y, Foriers A, Rogiers V, Vercruysse A
Dienst Toxicologie, Vrije Universiteit Brussel, Belgium.
Biochem Pharmacol. 1990 Feb 15;39(4):685-90. doi: 10.1016/0006-2952(90)90146-c.
Glutathione S-transferase (GST) isoenzymes of conventionally and co-cultured adult rat hepatocytes were purified and the GST subunits were separated by reversed phase HPLC in order to study the development of the GST subunit composition as a function of culture time and culture conditions. Several media conditions were tested, namely medium with and without fetal calf serum and with nicotinamide or dimethyl sulphoxide. Compared to the GST subunit composition of freshly isolated hepatocytes, changes in culture and media conditions result in a modification of the subunit profile. General observations are a decrease of subunits 1 and 2, an increase of subunit 3, a stabilization of subunit 4 and "de novo" expression of subunit 7. When [35S] methionine was added to the various culture media, and the thus labelled subunits were purified and separated, it was shown that cultured adult rat hepatocytes are able to synthesize the different GST proteins. Furthermore, the GST subunit composition, measured during various culture conditions, is probably a reflection of the "de novo" synthesis in vitro.
为了研究谷胱甘肽S-转移酶（GST）亚基组成随培养时间和培养条件的变化情况，对常规培养和共培养的成年大鼠肝细胞的GST同工酶进行了纯化，并通过反相高效液相色谱法分离了GST亚基。测试了几种培养基条件，即添加和不添加胎牛血清以及添加烟酰胺或二甲基亚砜的培养基。与新鲜分离的肝细胞的GST亚基组成相比，培养条件和培养基条件的变化导致亚基谱的改变。一般观察结果是亚基1和2减少，亚基3增加，亚基4稳定，亚基7“从头”表达。当向各种培养基中添加[35S]甲硫氨酸，并对标记的亚基进行纯化和分离时，结果表明培养的成年大鼠肝细胞能够合成不同的GST蛋白。此外，在各种培养条件下测得的GST亚基组成可能反映了体外的“从头”合成。
