期刊
MOLECULAR BIOTECHNOLOGY
卷 54, 期 2, 页码 250-256出版社
HUMANA PRESS INC
DOI: 10.1007/s12033-012-9562-3
关键词
Mucin; MUC2; MUC5AC; Recombinant protein; Chinese hamster ovary cells; Disulfide bonds; O-glycosylation
资金
- Swedish Research Council [7461, 21027, 342-2004-4434]
- Swedish Cancer Foundation
- Knut and Alice Wallenberg Foundation [KAW2007.0118]
- IngaBritt
- Arne Lundberg Foundation
- Sahlgren's University Hospital (LUA-ALF)
- Wilhelm and Martina Lundgren's Foundation
- Torsten och Ragnar Soderbergs Stiftelser
- Sahlgrenska Academy
- Swedish Foundation for Strategic Research-The Mucus-Bacteria-Colitis Center (MBC) of the Innate Immunity Program
The gel-forming mucins are large and heavily O-glycosylated proteins which build up mucus gels. The recombinant production of full-length gel-forming mucins has not been possible to date. In order to study mucin biosynthesis and biochemistry, we and others have taken the alternative approach of constructing different recombinant proteins consisting of one or several domains of these large proteins and expressing them separately in different cell lines. Using this approach, we have determined that MUC2, the intestinal gel-forming mucin, dimerizes via its C-terminal cysteine-knot domain and also trimerizes via one of the N-terminal von Willebrand D domains. Both of these interactions are disulfide bond mediated. Via this assembly, a molecular network is built by which the mucus gel is formed. Here we discuss not only the functional understanding obtained from studies of the recombinant proteins, but also highlight the difficulties encountered when these proteins were produced recombinantly. We often found an accumulation of the proteins in the ER and consequently no secretion. This was especially apparent when the cysteine-rich domains of the N- and C-terminal parts of the mucins were expressed. Other proteins that we constructed were either not secreted or not expressed at all. Despite these problems, the knowledge of mucin biosynthesis and assembly has advanced considerably through the studies of these recombinant proteins.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据