title: Quantitative analysis of aberrant protein glycosylation in liver cancer plasma by AAL-enrichment and MRM mass spectrometry.
abstract: A lectin-coupled mass spectrometry (MS) approach was employed to quantitatively monitor aberrant protein glycosylation in liver cancer plasma. To do this, we compared the difference in the total protein abundance of a target glycoprotein between hepatocellular carcinoma (HCC) plasmas and hepatitis B virus (HBV) plasmas, as well as the difference in lectin-specific protein glycoform abundance of the target glycoprotein. Capturing the lectin-specific protein glycoforms from a plasma sample was accomplished by using a fucose-specific aleuria aurantia lectin (AAL) immobilized onto magnetic beads via a biotin-streptavidin conjugate. Following tryptic digestion of both the total plasma and its AAL-captured fraction of each HCC and HBV sample, targeted proteomic mass spectrometry was conducted quantitatively by a multiple reaction monitoring (MRM) technique. From the MRM-based analysis of the total plasmas and AAL-captured fractions, differences between HCC and HBV plasma groups in fucosylated glycoform levels of target glycoproteins were confirmed to arise from both the change in the total protein abundance of the target proteins and the change incurred by aberrant fucosylation on target glycoproteins in HCC plasma, even when no significant change occurs in the total protein abundance level. Combining the MRM-based analysis method with the lectin-capturing technique proved to be a successful means of quantitatively investigating aberrant protein glycosylation in cancer plasma samples. Additionally, it was elucidated that the differences between HCC and control groups in fucosylated biomarker candidates A1AT and FETUA mainly originated from an increase in fucosylation levels on these target glycoproteins, rather than an increase in the total protein abundance of the target glycoproteins.
authors.affiliation: Division of Mass Spectrometry, Korea Basic Science Institute, 804-1 Yangcheong-Ri, Ochang-Eup, Cheongwon-Gun 363-883, Republic of Korea. jongshin@kbsi.re.kr.
authors.forename: Yeong Hee
authors.initials: YH
authors.lastname: Ahn
authors.forename: Park Min
authors.initials: PM
authors.lastname: Shin
authors.forename: Yong-Sam
authors.initials: YS
authors.lastname: Kim
authors.forename: Na Ree
authors.initials: NR
authors.lastname: Oh
authors.forename: Eun Sun
authors.initials: ES
authors.lastname: Ji
authors.forename: Kwang Hoe
authors.initials: KH
authors.lastname: Kim
authors.forename: Yeon Jung
authors.initials: YJ
authors.lastname: Lee
authors.forename: Sung Ho
authors.initials: SH
authors.lastname: Kim
authors.forename: Jong Shin
authors.initials: JS
authors.lastname: Yoo
mesh_terms: D015415:Biomarkers / Q000097:blood; D014408:Biomarkers, Tumor / Q000097:blood*; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D008113:Liver Neoplasms / Q000097:blood*; D013058:Mass Spectrometry / Q000379:methods*; D010949:Plasma / Q000737:chemistry / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D014408:Biomarkers, Tumor; D037102:Lectins; C064549:lectin, Aleuria aurantia