Differential proteomic approach for identification and verification of aberrantly glycosylated proteins in adenocarcinoma lung cancer (ADLC) plasmas by lectin-capturing and targeted mass spectrometry

Cited 12 time in scopus
Metadata Downloads
Title
Differential proteomic approach for identification and verification of aberrantly glycosylated proteins in adenocarcinoma lung cancer (ADLC) plasmas by lectin-capturing and targeted mass spectrometry
Author(s)
Y H Ahn; E S Ji; N R Oh; Yong Sam KimJeong Heon Ko; J S Yoo
Bibliographic Citation
Journal of Proteomics, vol. 106, pp. 221-229
Publication Year
2014
Abstract
To investigate quantitative differences in aberrant glycosylation of target glycoproteins between noncancerous group and patient group with adenocarcinoma lung cancer (ADLC), differential proteomic approach was developed by cooperatively using comparative lectin-capturing, targeted mass spectrometry (MRM MS), and antibody/lectin sandwich ELISA. Plasma samples comparatively prepared from 3 ADLC patients and 3 controls, with and without lectin-fractionation using fucose-specific Aleuria aurantia lectin (AAL), were trypsin-digested and analyzed for target glycoproteins, alpha-1-acid glycoprotein (AGP) and ceruloplasmin (CP), by MRM MS. From the MRM MS data the abundance levels of AAL-captured glycoforms of both targets were significantly higher in ADLC cases compared to controls, although the levels in total protein abundance were comparable between ADLC and control groups. This difference between ADLC and control groups in the fucosylated glycoform levels was originated mainly from aberrant fucosylation on the targets in ADLC plasmas rather than change in total protein abundance of the targets, and also confirmed by sandwich ELISA. AGP and CP were further verified to be biomarker candidates by MRM-based analysis of AAL-captured plasmas (30 ADLC cases, 30 controls), with AUROC 0.758 and 0.847 respectively. This differential proteomic approach can be useful for identifying and verifying biomarker candidate involved in aberrant protein glycosylation. Biological significance: The present paper introduces an efficient differential proteomic method to investigate quantitative differences in aberrant protein glycosylation of serological glycoproteins between noncancerous group and lung cancer patient group. This differential proteomic approach consisting of the targeted MRM MS of comparatively lectin-captured plasma fractions and the antibody/lectin sandwich ELISA-based assay was evaluated to be useful for identification of aberrantly fucosylated glycoproteins AGP and CP in lung cancer plasmas. In addition, we have demonstrated that the MRM MS-based differential proteomic approach is also useful for high-throughput verification of the aberrantly fucosylated glycoproteins AGP and CP using the large number of individual plasmas. Therefore, the present MRM MS-based differential proteomic strategy with lectin-capturing can be a powerful tool for high-throughput verification of aberrantly glycosylated biomarker candidates, identified preliminary by mass profiling experiments in proteomic fields but requiring further validation using a large number of cohorts.
Keyword
Aberrant glycosylationAdenocarcinoma lung cancerDifferential proteomicFucosylationTargeted mass spectrometry
ISSN
1874-3919
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.jprot.2014.04.031
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Genome Editing Research Center > 1. Journal Articles
Files in This Item:
  • There are no files associated with this item.


Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.