GLYCAN STRUCTURE & SITE OF GLYCOSYLATION IN RECOMBINANT ALPHA1 ANTITRYPSIN

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本研究的目的是确定重组α 1抗胰蛋白酶，<$1PI上的聚糖结构和糖基化位点的位置。 在本研究中，A1 PI在尼日尔曲霉中表达，并加载在SDS-PAGE和考马斯亮蓝染色。 从凝胶中切下两个主要条带，并分别研究每个级分（条带）的聚糖结构。 通过HPAEC、MALDI/TOF-MS、NSI-MSn和GC-MS分析释放的聚糖。我们发现，在两个库中，重组α 1 PI含有被几个末端半乳糖呋喃糖取代的高甘露糖型N-聚糖的异质混合物，但比例不同。 池B含有较高比例的较长聚糖，而池D含有较高比例的较短聚糖。 2008年，我们研究了a1 PI蛋白的N-糖基化位点定位。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The aim of the present study is to determine the glycan structure and the positions of glycosylation site(s) on the recombinant alpha1 antitrypsin, ¿1PI. In this study, A1PI was expressed in Aspergillus niger and loaded in SDS-PAGE and stained with Coomassie Blue. The two main bands were excised from the gel, and the glycan structure of each fraction (band) was investigated separately. The released glycans were analyzed by HPAEC, MALDI/TOF-MS, NSI-MSn, and GC-MS. We found that the recombinant a1PI, in both pools, contained heterogeneous mixture of high-mannose type N-glycans substituted with several terminal Galactofuranoses but the proportions were different. Pool B contains higher ratio of the longer glycans, whereas pool D contained higher portions of shorter glycans. In 2008, we studied the N-glycosylation site mapping of a1PI protein.