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    • 89. 发明申请
    • CO-EXPRESSION OF PROLINE HYDROXYLASES TO FACILITATE HYP-GLYCOSYLATION OF PROTEINS EXPRESSED AND SECRETED IN PLANT CELLS
    • 共表达丙氨酸羟基来促进植物细胞中表达和分泌的蛋白质的高糖蛋白化
    • WO2008008766A3
    • 2008-11-20
    • PCT/US2007073137
    • 2007-07-10
    • UNIV OHIOKIELISZEWSKI MARCIAXU JIANFENGCOOPER IVER P
    • KIELISZEWSKI MARCIAXU JIANFENGCOOPER IVER P
    • C12N15/82
    • C12N15/8257C07K14/78C12N9/0071C12P21/005
    • Plant cells contain particular proline hydroxylases whose specificity determines which prolines, in a protein expressed therein, are hydroxylated. The resulting hydroxyprolines, in turn, my be glycosylated. It is the presence of glycosylated hydroxyprolines which is the most important determinant of the degree of secretion of the protein. One may modify the proline hydroxylation capabilities of the plant cell by co-expressing in that cell at least one exogeneous proline hyrdoxylase which will hydroxylate at least one proline site in the protein which is not hydroxylated by the endogeneous proline hydroxylase, but which, if hydroxylated, can than be glycosylated by the plant cell, resulting in increased Hyp-glycosylation. Moreover, even if the co-expressed exogenous proline hydroxylase doesn't achieve the desired level of Hyp-glycosylation in the native protein, such co-expression may still be used in conjunction with modification of the protein itself to increase the number and/or the likelihood of Hyp-glycosylation sites.
    • 植物细胞含有特定的脯氨酸羟化酶,其特异性决定了其中表达的蛋白质中的哪些脯氨酸被羟基化。 所得的羟基脯氨酸又被我糖基化。 糖化羟脯氨酸的存在是蛋白质分泌程度最重要的决定因素。 人们可以通过在该细胞中共表达至少一种外源脯氨酸脱乙酰酶,来改变植物细胞的脯氨酸羟基化能力,该外源脯氨酸脱乙酰酶能使蛋白质中的至少一个脯氨酸位点羟基化而不被内源脯氨酸羟化酶羟基化,但如果羟基化 ,可以被植物细胞糖基化,导致Hyp-糖基化的增加。 此外,即使共表达的外源脯氨酸羟化酶在天然蛋白质中没有达到所需的Hyp糖基化水平,这种共表达仍然可以与蛋白质本身的修饰结合使用以增加数量和/或 低糖基化位点的可能性。