摘要
根据枸橼酸酐对蛋白质中的游离氨基进行化学修饰后可使蛋白质溶解度提高的原理，以大肠杆菌表达的人重组ＧＭ－ＣＳＦ为模型，研究了枸橼酸酐修饰对含凝血酶识别位点的融合蛋白的作用，发现用微量枸橼酸酐修饰的重组ＧＭ－ＣＳＦ变性、复性更容易，溶解度明显提高，并对凝血酶的消化更为敏感，使凝血酶用量降低１００倍．ＧＭ－ＣＳＦ活性测定结果证明枸橼酸酐修饰不影响其生物学活性．这些结果为枸橼酸酐修饰法在大肠杆菌表达重组蛋白纯化中的应用提供了新途径．
The solubility of the protein can beincreased by citraconic anhydride(CT)modification. According to this principle; the effects ofthe CT modification on the fusion protein usingrhGM-CSF as a model which contain thrombincleavage sites have been studied.The resultdemonstrated that the process of denature andrenature in the modified protein was much easierthan that in unmodified counterpart. In additionthe modified protein is more sensitive to thrombin digestion; one percent amount of thrombinwas enough to achieve the complete digestion.The modification by CT did not effect the bioactivity of rhGM-CSF.A new way was paved inthe purification of recombinant protein by CTmodification method.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1996年第6期541-544,共4页
Progress In Biochemistry and Biophysics
