Heikoop J C, Grootenhuis P D, Blaauw M, Veldema J S, Van Haastert P J, Linskens M H
N.V. Organon, Oss, The Netherlands.
Eur J Biochem. 1998 Sep 1;256(2):359-63. doi: 10.1046/j.1432-1327.1998.2560359.x.
Human choriogonadotropin (hCG) is a highly complex glycoprotein consisting of two non-covalently associated subunits. We aimed for the expression of a single-chain hCG in the soil amoebae Dictyostelium discoideum, a host which, in principle, provides simple genetics in combination with complex protein synthesis. To limit anticipated problems in mRNA translation, the first 30 bases of the coding sequence were altered to conform to the Dictyostelium preferred codon usage. We show that, immunologically, active single-chain hCG is indeed produced by Dictyostelium. Furthermore, this single-chain hCG is able to bind to the human luteinizing hormone/CG receptor and elicit a biological response. Its receptor-binding affinity is comparable to single-chain hCG produced by mammalian cells. We conclude that Dictyostelium is able to express bioactive highly complex heterologous glycoproteins.
人绒毛膜促性腺激素（hCG）是一种高度复杂的糖蛋白，由两个非共价结合的亚基组成。我们旨在在土壤变形虫盘基网柄菌中表达单链hCG，该宿主原则上能提供简单的遗传学方法以及复杂的蛋白质合成机制。为了限制mRNA翻译中预期出现的问题，编码序列的前30个碱基被改变，以符合盘基网柄菌偏好的密码子使用方式。我们发现，从免疫学角度来看，盘基网柄菌确实产生了具有活性的单链hCG。此外，这种单链hCG能够与人促黄体生成素/CG受体结合并引发生物学反应。其受体结合亲和力与哺乳动物细胞产生的单链hCG相当。我们得出结论，盘基网柄菌能够表达具有生物活性的高度复杂的异源糖蛋白。
