fulltext.study @t Gmail

Binding activity of recombinant human L-selectin-Fcγ is modified by sialylation

Paper ID Volume ID Publish Year Pages File Format Full-Text
4042 205 2010 7 PDF Available
Title
Binding activity of recombinant human L-selectin-Fcγ is modified by sialylation
Abstract

The adhesion molecule L-selectin expressed on most leukocytes mediates tethering and rolling of leukocytes on activated endothelia and initiates the extravasation of leukocytes into inflamed tissues. Recombinant L-selectin-Fcγ is widely used both as a tool to study this key step of inflammation and as an anti-inflammatory compound in animal models of inflammation. Since previous studies on cellular L-selectin have indicated that glycosylation influences adhesive interactions of the adhesion molecule, we have examined whether the binding activity of L-selectin-Fcγ is affected by sialylation. Different forms of recombinant human L-selectin-Fcγ were expressed in CHO and K-562 cells and were purified by affinity chromatography using Protein A-Sepharose. A hypersialylated form of L-selectin-Fcγ was generated by culturing cells in the presence of 5 mM N-acetyl-beta-d-mannosamine, while a desialylated variant was obtained by treatment of purified L-selectin-Fcγ with neuraminidase. Binding activity to the synthetic biligand SiaLex-PAA-sTyr was measured by surface plasmon resonance (SPR) technology. While hypersialylated L-selectin-Fcγ showed decreased binding activity, desialylation elevated L-selectin-Fcγ binding to SiaLex-PAA-sTyr. The data show that sialylation of L-selectin-Fcγ reduces binding activity to ligand epitopes containing sialyl Lewis x and sulfated tyrosine residues. For the production of biologically active L-selectin-Fcγ conditions should be chosen that favor the generation of non-sialylated or of scarcely sialylated forms of the recombinant glycoprotein.

Keywords
L-selectin; Neuraminidase; Recombinant protein production; Sialic acid; Sialylation; Surface plasmon resonance
First Page Preview
Binding activity of recombinant human L-selectin-Fcγ is modified by sialylation
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 48, Issue 2, 15 January 2010, Pages 253–259
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us