Marlovits T C, Zechmeister T, Gruenberger M, Ronacher B, Schwihla H, Blaas D
Institute of Biochemistry, University of Vienna, Austria.
FASEB J. 1998 Jun;12(9):695-703. doi: 10.1096/fasebj.12.9.695.
A fragment of the low density lipoprotein receptor encompassing the seven ligand binding repeats was expressed in Sf9 insect cells as a fusion protein with a carboxyl-terminally linked hexa-his tag by using a baculovirus vector. Up to 10 mg/l of the fusion protein was secreted into the medium. The material was soluble in the absence of detergent and active in binding beta very low density lipoprotein and a member of the minor group of human rhinoviruses (HRV2) in ligand blots from sodium dodecyl sulfate-polyacrylamide gels run under nonreducing conditions. The receptor fragment specifically inhibits viral infection of HeLa cells by minor group HRVs in a concentration-dependent manner. Viral infectivity is neutralized by aggregation.
利用杆状病毒载体，将包含七个配体结合重复序列的低密度脂蛋白受体片段作为与羧基末端连接的六聚组氨酸标签的融合蛋白，在Sf9昆虫细胞中表达。高达10mg/l的融合蛋白分泌到培养基中。该物质在没有去污剂的情况下可溶，并且在非还原条件下运行的十二烷基硫酸钠-聚丙烯酰胺凝胶的配体印迹中，能与β极低密度脂蛋白和一小类人类鼻病毒（HRV2）的成员结合。该受体片段以浓度依赖的方式特异性抑制HeLa细胞被小类HRV的病毒感染。病毒感染性通过聚集而被中和。
