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Title: Expression, purification and characterization of C2 domain of milk fat globule-EGF-factor 8-L
Authors: Nanga, Ravi Prakash Reddy
Vivekanandan, Subramanian
Yoon, Ho Sup
Keywords: DRNTU::Science::Biological sciences
Issue Date: 2006
Source: Nanga, R. P. R., Vivekanandan, S., & Yoon, H. S. (2007). Expression, purification and characterization of C2 domain of milk fat globule-EGF-factor 8-L. Protein Expression and Purification, 52(2), 329-333.
Series/Report no.: Protein expression and purification
Abstract: Milk fat globule-EGF-factor 8-L (MFG-E8L) is secreted by activated macrophages and functions as a linker protein or opsonin between the dying cells and phagocytes. MFG-E8L recognizes the apoptotic or dying cells by specifically binding to Phosphatidylserine (PS) exposed on the outer cell surface and enhances the engulfment of the apoptotic cells by phagocytes, thereby preventing the inflam¬mation and autoimmune response against intracellular antigens that can be released from the dying cells. MFG-E8L contains two EGF¬like domains, P/T (proline/threonine) rich domain followed by two discoidin-like domains (C1 and C2). Recent studies have shown that the C2 domain of MFG-E8L is specifically involved in interaction with PS exposed on the apoptotic cells. Towards understanding this specific molecular interaction between the MFG-E8L C2 domain and PS, we expressed, purified the C2 domain of MFG-E8L and per¬formed the binding studies with phospholipids by 31P NMR experiment. We demonstrated that our recombinant construct and expres¬sion system were effective and allowed us to obtain the C2 domain and also showed that the purified C2 domain was stable and properly folded, and our 31P NMR studies indicated that the C2 domain had specific binding with PS.
ISSN: 10465928
DOI: 10.1016/j.pep.2006.08.018
Schools: School of Biological Sciences 
Rights: © 2006 Elsevier. This is the author created version of a work that has been peer reviewed and accepted for publication by Protein Expression and PuriWcation, Elsevier. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at:
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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