Li Hua, Chavan Manasi, Schindelin Hermann, Lennarz William J, Li Huilin
Biology Department, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
Structure. 2008 Mar;16(3):432-40. doi: 10.1016/j.str.2007.12.013.
Oligosaccharyl transferase (OT) catalyzes the transfer of a lipid-linked oligosaccharide to the nascent polypeptide emerging from the translocon. Currently, there is no structural information on the membrane-embedded OT complex, which consists of eight different polypeptide chains. We report a 12 A resolution cryo-electron microscopy structure of OT from yeast. We mapped the locations of four essential OT subunits through a maltose-binding protein fusion strategy. OT was found to have a large domain in the lumenal side of endoplasmic reticulum where the catalysis occurs. The lumenal domain mainly comprises the catalytic Stt3p, the donor substrate-recognizing Wbp1p, and the acceptor substrate-recognizing Ost1p. A prominent groove was observed between these subunits, and we propose that the nascent polypeptide from the translocon threads through this groove while being scanned by the Ost1p subunit for the presence of the glycosylation sequon.
寡糖基转移酶(OT)催化脂质连接的寡糖转移到从转运体中出现的新生多肽上。目前,尚无关于由八条不同多肽链组成的膜嵌入OT复合物的结构信息。我们报道了酵母OT的12埃分辨率冷冻电子显微镜结构。我们通过麦芽糖结合蛋白融合策略确定了四个必需OT亚基的位置。发现OT在内质网腔侧有一个大结构域,催化作用在此发生。腔结构域主要由催化性的Stt3p、识别供体底物的Wbp1p和识别受体底物的Ost1p组成。在这些亚基之间观察到一个明显的凹槽,我们推测来自转运体的新生多肽穿过这个凹槽,同时被Ost1p亚基扫描以检测糖基化序列的存在。