AAV8 衣壳蛋白上的位点特异性 N-糖基化。
Site-Specific N-Glycosylation on the AAV8 Capsid Protein.
机构信息
Center for Diagnostics & Therapeutics and Department of Chemistry, Georgia State University, Atlanta, GA 30302, USA.
Sol Sherry Thrombosis Research Center, Lewis Katz, School of Medicine, Temple University, Philadelphia, PA 19140, USA.
出版信息
Viruses. 2018 Nov 17;10(11):644. doi: 10.3390/v10110644.
Adeno associated virus (AAV) is a versatile gene delivery tool, which has been approved as a human gene therapy vector for combating genetic diseases. AAV capsid proteins are the major components that determine the tissue specificity, immunogenicity and in vivo transduction performance of the vector. In this study, the AAV8 capsid glycosylation profile was systemically analyzed by peptide mass fingerprinting utilizing high-resolution mass spectrometry to determine the presence of capsid glycosylation. We identified N-glycosylation on the amino acid N499 of the capsid protein. We characterized the overall sugar profile for vector produced in 293 cells. Multiple N-glycosylated host-cell proteins (HCPs) copurified with AAV8 vectors and were identified by analyzing LC-MS data utilizing a human database and proteome discoverer search engine. The N-glycosylation analysis by MALDI-TOF MS, highlighted the probability of AAV8 interaction with terminal galactosylated N-glycans within the HCPs.
腺相关病毒（AAV）是一种多功能的基因传递工具，已被批准作为治疗遗传疾病的人类基因治疗载体。AAV 衣壳蛋白是决定载体组织特异性、免疫原性和体内转导性能的主要成分。在这项研究中，我们通过利用高分辨率质谱进行肽质量指纹图谱分析，系统地分析了 AAV8 衣壳的糖基化谱，以确定衣壳糖基化的存在。我们在衣壳蛋白的氨基酸 N499 上鉴定出 N-糖基化。我们对在 293 细胞中产生的载体的整体糖谱进行了表征。通过分析利用人类数据库和 proteome discoverer 搜索引擎的 LC-MS 数据，鉴定出与 AAV8 载体共纯化的多种 N-糖基化宿主细胞蛋白（HCP）。MALDI-TOF MS 的 N-糖基化分析强调了 AAV8 与 HCP 中末端半乳糖化 N-聚糖相互作用的可能性。
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