Comprehensive glyco-proteomic analysis of human alpha1-antitrypsin and its charge isoforms

Proteomics
Daniel KolarichFriedrich Altmann

Abstract

Human alpha1-antitrypsin (A1PI) is a well-known glycoprotein in human plasma important for the protection of tissues from proteolytic enzymes. The three N-glycosylation sites of A1PI contain diantennary N-glycans but also triantennary and even traces of tetraantennary structures leading to the typical IEF pattern observed for A1PI. Here we present an approach to characterize A1PI isoforms from human plasma and its PTMs by LC-ESI-MS and LC-ESI-MS/MS of peptides obtained by proteolytic digestion. The single cysteine residue of A1PI formed a disulfide bridge with free cysteine. The variability of the number of antennae and hence sialic acids on glycosylation site N107, which even contained minute amounts of tetraantennary structures, emerged as a major cause for the IEF pattern of A1PI. Only negligible amounts of triantennary structures were identified attached to N70, and exclusively diantennary structures were present on site N271 in each of the isoforms analyzed. Exoglycosidase digests revealed alpha2,6-linked neuraminic acids on diantennary N-glycans, and triantennary contained additionally one single alpha2,3-neuraminic acid per N-glycan, which, together with a fucose, formed a sialyl Lewis X determinant on the beta1,4-linked...Continue Reading

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Citations

Feb 22, 2012·Archivum Immunologiae Et Therapiae Experimentalis·David A BerginEmer P Reeves
Apr 5, 2011·Analytical and Bioanalytical Chemistry·Josephine GrassFriedrich Altmann
Feb 22, 2012·Applied Microbiology and Biotechnology·Martin PfefferDiethard Mattanovich
Jul 22, 2008·Glycoconjugate Journal·François FenailleNicolas Bihoreau
Aug 14, 2012·Glycoconjugate Journal·Jonas NilssonGöran Larson
Oct 16, 2012·Glycoconjugate Journal·Kyung Jin LeeDoo-Byoung Oh
Apr 23, 2013·Journal of the American Society for Mass Spectrometry·Arun V Everest-DassMatthew P Campbell
Oct 5, 2013·Journal of Proteome Research·Benjamin L ParkerStuart J Cordwell
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Aug 19, 2009·Proteomics·Johannes StadlmannFriedrich Altmann
Oct 2, 2008·Biotechnology and Bioengineering·Liat ChillJoseph Shiloach
Apr 20, 2007·Proteomics·Daniel KolarichFriedrich Altmann
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Jan 19, 2008·Proteomics·Daniel KolarichFriedrich Altmann
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Jun 15, 2013·Electrophoresis·Weijie DongAkihiko Kameyama

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