https://scholars.lib.ntu.edu.tw/handle/123456789/630083
標題: | Site-Specific N-Glycosylation of Caprine Lysostaphin Restricts its Bacteriolytic Activity Toward Staphylococcus Aureus | 作者: | Huang, Ching Ying JIH-TAY HSU Chung, Pei Hsuan Cheng, Winston Teng Kuei Jiang, Yan Nian YU-TEN JU |
關鍵字: | Glycosylation | Goat | Lysostaphin | Mastitis | 公開日期: | 1-四月-2013 | 出版社: | TAYLOR & FRANCIS INC | 卷: | 24 | 期: | 2 | 起(迄)頁: | 129 | 來源出版物: | Animal Biotechnology | 摘要: | Lysostaphin (LYS) is an anti-staphylococcal prokaryotic polypeptide that has been used to avoid Staphylococcus aureus mastitis through transgenic or viral vector approaches exogenously expressed in dairy animals. However, glycosylation of lysostaphin expressed in mammalian cells results in a loss of bioactivity. Until now, the mechanism of site-specific glycosylation of lysostaphin causing this loss of bioactivity remains unknown. An immortalized caprine mammary epithelial cell line (CMEC-08-D) was used to study recombinant lysostaphin fused with goat β-casein, goat lactoferrin (LF) or prokaryotic signal peptides. These constructs were separately ectopically expressed in CMEC-08-D. Results of site-directed mutagenesis show that Asn125 but not Asn232 is the exact glycosylation site of lysostaphin expressed in CMEC-08-D. In addition, the effect of glycosylation of lysostaphin on its staphylolytic activity was identified through bacterial plate assay. The data indicated that wild type and mutated N232Q-lysostaphin (Asn232 to Gln232 substitution) lacked staphylolytic activity. In contrast, mutated N125Q (Asn125 to Gln125 substitution) and N125Q/N232Q-lysostaphin possessed staphylolytic activity. On the other hand, all mutated lysostaphin showed no change in binding ability to S. aureus. This reveals that N-glycosylation at Asn125 of lysostaphin expressed in a eukaryotic system greatly decreases lysostaphin bacteriolytic activity but does not affect its binding ability to S. aureus. © 2013 Copyright Taylor and Francis Group, LLC. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-84880940461&doi=10.1080%2f10495398.2012.760469&partnerID=40&md5=1adffe24b709c5c1e25268f7e3df2107 https://scholars.lib.ntu.edu.tw/handle/123456789/630083 |
ISSN: | 10495398 | DOI: | 10.1080/10495398.2012.760469 |
顯示於: | 動物科學技術學系 |
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