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- W2610974860 abstract "Anterior gradient protein 2 homolog is a member ofprotein-disulfide isomerase (PDI) family expressed inendoplasmatic reticulum. The suggested function is aco-chaperone aiding the forming of disulfide bonds with role intumour metastasis. Little knowledge is available aboutAGR2-interacting or –binding partners in cells. To date, onlyMucin 2 was described as the disulfide-linked AGR2 partner andthree other proteins reptin, LYPD3 and DAG1 were predicted tointeract with AGR2 by two-hybrid yeast study. We analysedpossible AGR2 partners using cell line with endogenous AGR2expression (T47D) as well as AGR2-stable transfected cells(H1299). We applied reversible protein cross-linking (DSPagent) on cells in vivo to stabilize protein complexes andprecipitated AGR2 with AGR2 specific and un-specific peptidesand antibodies followed by Filter Aided Proteome preparation(FASP) and high resolution mass spectrometry analysis of IPeluates (LC-MS/MS on Orbitrap instrument). Label-FreeQuantification (LFQ) of proteins identified inSwissprot/Uniprot database was performed in MaxQuant dataanalysis software. Applying described method with eliminationof un-specifically bound proteins we obtained AGR2 partners inAGR2-specific IP eluates. PDI members A3 and A6 were repeatedlyidentified in both cell lines, while AGR3 protein was foundonly in T47D cells based on different expression in cell lines,quantitatively the proteins are significantly presented inhigher levels in cross-linked samples in comparison toun-cross-linked control." @default.
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- W2610974860 date "2012-09-24" @default.
- W2610974860 modified "2023-09-27" @default.
- W2610974860 title "Identification of AGR2 partners using mass spectrometry withlabel-free quantification" @default.
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