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. 2015 Mar 11;17(1):36. doi: 10.1186/s13058-015-0544-9

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© Zhou et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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The identification of RCA-I-recognized membrane glycoprotein by SILAC and LC-MS/MS analysis. (a) A SILAC and LC-MS/MS combined strategy was used to detect both expression levels and galactosylations of the membrane glycoproteins captured by RCA-I. (b) LC-MS/MS spectrum of a peptide. The sequence of the peptide was identified as VAPEEHPVLLTEAPLNPK, which was from POTEF. (c) Representative base peak chromatogram of POTEF in the whole-membrane fraction (top) and RCA-I-enriched membrane fraction (down). The blue lines represent peptides of POTEF from 4175 cells whereas the red ones represent those from SCP2 cells (¹³C-labeled). LC-MS/MS, liquid chromatography-mass spectrometry/tandem spectrometry; POTEF, POTE ankyrin domain family member F; RCA-I, Ricinus communis agglutinin I; SILAC, stable isotope labeling by amino acids in cell culture.