Gill G S, Zaworski P G, Marotti K R, Rehberg E F
Molecular Biology Research, Upjohn Company, Kalamazoo, MI 49007.
Biotechnology (N Y). 1990 Oct;8(10):956-8. doi: 10.1038/nbt1090-956.
We have developed a simple screening procedure that allowed us to identify Saccharomyces cerevisiae strains able to secrete human tissue plasminogen activator (tPA) into the culture medium. The screen can be used to isolate more efficient secretor strains and to look for novel tPA analogs. Employing one of these strains to study the effect of glycosylation on secretion, we show that glycosylation in the catalytic domain of tPA plays an important role in folding and/or secretion of the molecule. Removing this glycosylation site resulted in a 3-5-fold reduction in the level of tPA secretion. We anticipate that this system will prove useful in studying yeast secretory pathway as well as structure-function relationships in the tPA molecule.
我们开发了一种简单的筛选程序，可用于鉴定能够将人组织型纤溶酶原激活剂（tPA）分泌到培养基中的酿酒酵母菌株。该筛选方法可用于分离更高效的分泌菌株，并寻找新型tPA类似物。利用其中一种菌株研究糖基化对分泌的影响，我们发现tPA催化结构域中的糖基化在分子折叠和/或分泌中起重要作用。去除该糖基化位点导致tPA分泌水平降低3至5倍。我们预计该系统将被证明在研究酵母分泌途径以及tPA分子的结构-功能关系方面很有用。
