Gahmberg C G, Hakomori S I
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3329-33. doi: 10.1073/pnas.70.12.3329.
By use of galactose oxidase (EC 1.1.3.9), followed by reduction with tritiated sodium borohydride, the surface structures of transformed 3T3 and NIL cells, under ordinary growth conditions, were characterized by (i) deletion of the normally existing glycoprotein label and (ii) appearance or increase of a new glycoprotein label. NIL cells had a galactoprotein label with molecular weight 200,000 that was deleted in NIL cells transformed by polyoma virus. 3T3 cells had a glycoprotein label with molecular weight of 30,000 that was lost after transformation. Glycoproteins of transformed 3T3 cells, with molecular weight 105,000, and those of transformed NIL cells, with molecular weight 85,000, were not labeled in normal confluent cells, but became labeled after trypsin treatment. The label in glycolipids was quantitatively different in normal and transformed cells. The labeling pattern in glycoprotein and glycolipids of transformed NIL and 3T3 cells became similar to that of nontransformed cells when contact responses of transformed cells became conspicuous when cells were cultured in the presence of dextran sulfate or dibutyryl cyclic adenosine monophosphate, or in medium in which glucose was replaced with galactose.
通过使用半乳糖氧化酶(EC 1.1.3.9),随后用氚化硼氢化钠还原,在普通生长条件下,转化的3T3细胞和NIL细胞的表面结构具有以下特征:(i)正常存在的糖蛋白标记缺失,以及(ii)新的糖蛋白标记出现或增加。NIL细胞具有分子量为200,000的半乳糖蛋白标记,该标记在被多瘤病毒转化的NIL细胞中缺失。3T3细胞具有分子量为30,000的糖蛋白标记,该标记在转化后丢失。转化的3T3细胞中分子量为105,000的糖蛋白以及转化的NIL细胞中分子量为85,000的糖蛋白,在正常汇合细胞中未被标记,但在胰蛋白酶处理后被标记。正常细胞和转化细胞中糖脂的标记在数量上有所不同。当在硫酸葡聚糖或二丁酰环磷酸腺苷存在下培养细胞,或在葡萄糖被半乳糖替代的培养基中培养细胞时,转化的NIL细胞和3T3细胞的糖蛋白和糖脂标记模式变得与未转化细胞相似,此时转化细胞的接触反应变得明显。