Punt C J, Rijksen G, Vlug A M, van der Heijden M C, Dekker A W, Staal G E
Department of Medical Oncology, University Hospital Nijmegen, The Netherlands.
Leuk Res. 1991;15(12):1107-16. doi: 10.1016/0145-2126(91)90178-v.
We have examined the expression of the protein tyrosine kinase (PTK) encoding oncogenes fes and abl in normal and malignant human myeloid cells in immunoblotting experiments. fes was markedly present in all cytosolic and most membrane fractions of normal and malignant cells. abl was only visible in normal cells, and occurred mostly in the cytosolic fractions. Molecular weights of identified proteins were different from the known products of fes and abl, possibly by alternative splicing at the mRNA level or by proteolysis. PTKs in myeloid cells were further purified by fast liquid protein chromatography (FPLC). PTK-activities of column fractions were assayed using a solid-phase non-radioactive dot-blot assay. Cytosolic and membrane fractions showed a FPLC pattern with a constant as well as a variable part in both normal and malignant cells, possibly indicative for PTKs with specialized functions in normal cell growth and transformation. Partial characterization of PTKs from different eluted peaks of AML-M4 blast cells demonstrated that PTKs from these peaks are kinetically distinct from each other.
我们在免疫印迹实验中检测了编码癌基因fes和abl的蛋白酪氨酸激酶（PTK）在正常和恶性人髓细胞中的表达。fes明显存在于正常和恶性细胞的所有胞质部分以及大多数膜部分中。abl仅在正常细胞中可见，且主要存在于胞质部分。鉴定出的蛋白质的分子量与fes和abl的已知产物不同，这可能是由于mRNA水平的可变剪接或蛋白水解所致。通过快速液相蛋白色谱法（FPLC）进一步纯化髓细胞中的PTK。使用固相非放射性点杂交法测定柱馏分的PTK活性。胞质和膜部分在正常和恶性细胞中均显示出具有恒定部分和可变部分的FPLC图谱，这可能表明PTK在正常细胞生长和转化中具有特殊功能。对急性髓系白血病M4原始细胞不同洗脱峰的PTK进行的部分表征表明，这些峰的PTK在动力学上彼此不同。
