title: A multiplex lectin-channel monitoring method for human serum glycoproteins by quantitative mass spectrometry.
abstract: A mass profiling method and multiple reaction monitoring (MRM)-based quantitative approach were used to analyze multiple lectin-captured fractions of human serum using different lectins such as aleuria aurantia lectin (AAL), phytohemagglutinin-L(4) (L-PHA), concanavalin A (Con A), and Datura stramonium agglutinin (DSA) to quantitatively monitor protein glycosylation diversity. Each fraction, prepared by multiple lectin-fractionation and tryptic digestion, was analyzed by 1-D LC-MS/MS. Semi-quantitative profiling showed that the list of glycoproteins identified from each lectin-captured fraction is significantly different according to the used lectin. Thus, it was confirmed that the multiplex lectin-channel monitoring (LCM) using multiple lectins is useful for investigating protein glycosylation diversity in a proteome sample. Based on the semi-quantitative mass profiling, target proteins showing lectin-specificity among each lectin-captured fraction were selected and analyzed by the MRM-based method in triplicate using each lectin-captured fraction (average CV 7.9%). The MRM-based analysis for each lectin-captured fraction was similar to those obtained by the profiling experiments. The abundance of each target protein measured varied dramatically, based on the lectin-specificity. The multiplex LCM approach using MRM-based analyses is useful for quantitatively monitoring target protein glycoforms selectively fractionated by multiple lectins. Thus through multiplex LCM rather than single, we could inquire minutely into protein glycosylation states.
authors.affiliation: Division of Mass Spectrometry, Korea Basic Science Institute, 804-1 Yangcheong-Ri, Ochang-Myun, Cheongwon-Gun, 363-883, Republic of Korea.
authors.forename: Yeong Hee
authors.initials: YH
authors.lastname: Ahn
authors.forename: Eun Sun
authors.initials: ES
authors.lastname: Ji
authors.forename: Park Min
authors.initials: PM
authors.lastname: Shin
authors.forename: Kwang Hoe
authors.initials: KH
authors.lastname: Kim
authors.forename: Yong-Sam
authors.initials: YS
authors.lastname: Kim
authors.forename: Jeong Heon
authors.initials: JH
authors.lastname: Ko
authors.forename: Jong Shin
authors.initials: JS
authors.lastname: Yoo
mesh_terms: D001798:Blood Proteins / Q000032:analysis*; D002853:Chromatography, Liquid; D006023:Glycoproteins / Q000097:blood*; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001798:Blood Proteins; D006023:Glycoproteins; D037102:Lectins