Linked Life Data

21495009

Source:http://linkedlifedata.com/resource/pubmed/id/21495009

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
2011-7-19
pubmed:abstractText
Human alpha-1-antitrypsin (A1AT) is a protease inhibitor that is involved in the protection of lungs from neutrophil elastase enzyme that drastically modifies tissue functioning. The glycoprotein consists of 394 amino acids and is N-glycosylated at Asn-46, Asn-83, and Asn-247. A1AT deficiency is currently treated with A1AT that is purified from human serum. In view of therapeutic applications, rA1AT was produced using a novel human neuronal cell line (AGE1.HN®) and we investigated the N-glycosylation pattern as well as the in vitro anti-inflammatory activity of the recombinant glycoprotein. rA1AT (300 mg/L) was biologically active as analyzed using elastase assay. The N-glycan pool, released by PNGase F digestion, was characterized using 2D-HPLC, MALDI-TOF mass spectrometry, and by exoglycosidase digestions. A total of 28 N-glycan structures were identified, ranging from diantennary to tetraantennary complex-type N-glycans. Most of the N-glycans were found to be (?1-6) core-fucosylated and part of them contain the Lewis X epitope. The two major compounds are a monosialylated diantennary difucosylated glycan and a disialylated diantennary core-fucosylated glycan, representing 25% and 18% of the total N-glycan pool, respectively. Analysis of the site-specificity revealed that Asn-247 was mainly occupied by diantennary N-glycans whereas Asn-46 was occupied by di-, and triantennary N-glycans. Asn-83 was exclusively occupied by sialylated tri- and tetraantennary N-glycans. Next, we evaluated the anti-inflammatory activity of rA1AT using A1AT purified from human serum as a reference. rA1AT was found to inhibit the production of TNF-? in neutrophils and monocytes as commercial A1AT does.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
1097-0290
pubmed:author
pubmed:copyrightInfo
Copyright © 2011 Wiley Periodicals, Inc.
pubmed:issnType
Electronic
pubmed:volume
108
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2118-28
pubmed:meshHeading
pubmed-meshheading:21495009-Anti-Inflammatory Agents, pubmed-meshheading:21495009-Biotechnology, pubmed-meshheading:21495009-Cell Line, pubmed-meshheading:21495009-Cells, Cultured, pubmed-meshheading:21495009-Chromatography, High Pressure Liquid, pubmed-meshheading:21495009-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:21495009-Glycosylation, pubmed-meshheading:21495009-Humans, pubmed-meshheading:21495009-Leukocyte Elastase, pubmed-meshheading:21495009-Monocytes, pubmed-meshheading:21495009-Neurons, pubmed-meshheading:21495009-Neutrophils, pubmed-meshheading:21495009-Recombinant Proteins, pubmed-meshheading:21495009-Sialic Acids, pubmed-meshheading:21495009-Spectrometry, Mass, Matrix-Assisted Laser..., pubmed-meshheading:21495009-Tumor Necrosis Factor-alpha, pubmed-meshheading:21495009-alpha 1-Antitrypsin
pubmed:year
2011
pubmed:articleTitle
N-glycosylation and biological activity of recombinant human alpha1-antitrypsin expressed in a novel human neuronal cell line.
pubmed:affiliation
Glycodesign and Glycoanalytics, Central Institute of Laboratory Medicine and Pathobiochemistry, Charité Medical University, Charité Platz 1, 10117 Berlin, Germany. veronique.blanchard@charite.de
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't