Purification and identification of transferrin as a major pituitary-derived mitogen for MTW9/PL2 rat mammary tumor cells.
Purification and identification of transferrin as a major pituitary-derived mitogen for MTW9/PL2 rat mammary tumor cells.
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转铁蛋白的纯化和鉴定作为 MTW9/PL2 大鼠乳腺肿瘤细胞的主要垂体衍生有丝分裂原。
DOI：
10.1007/bf02620963
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Sirbasku,DA
中科院分区：
文献类型：
--
作者：
Riss,TL;Sirbasku,DA
Transferrin was identified as a major tissue-derived growth factor for MTW9/PL2 rat mammary tumor cells. Mitogenic activity was assayed by the ability to stimulate the increase in number of MTW9/PL2 cells over 4 d in Dulbecco's modified Eagle's medium containing only 15 mMHEPES, 2 mMglutamine, and 50 μg/ml gentamicin. This growth-promoting activity was purified from ammonium sulfate precipitates of phosphate buffered saline extracts of porcine pituitaries using DEAE-Sepharose, chromatofocusing, molecular sieve chromatography and reverse phase high performance liquid chromatography. Pig pituitary mitogen (PPM) migrated as a single band at molecular weight 78 000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis, eluted from chromatofocusing at multiple pH values near 6.3, exhibited an absorption maximum at 465 nm which was diminished by removal of iron, showed a characteristic salmon-pink color in aqueous solution, and was similar in amino acid composition to previously reported values for porcine transferrin. Purified PPM similar to commercially available human transferrin (ED50=160 to 350 ng/ml). We have concluded that using serum-free assay conditions with MTW9/PL2 cells, transferrin was a major source of the mitogenic activity present in extracts of porcine pituitary.
