From glycomics to functional glycomics of sugar chains: Identification of target proteins with functional changes using gene targeting mice and knock down cells of FUT8 as examples.
Comprehensive analyses of proteins from cells and tissues are the most effective means of elucidating the expression patterns of individual disease-related proteins. On the other hand, the simultaneous separation and characterization of proteins by 1-DE or 2-DE followed by MS analysis are one of the fundamental approaches to proteomic analysis. However, these analyses do not permit the complete structural identification of glycans in glycoproteins or their structural characterization. Over half of all known proteins are glycosylated and glycan analyses of glycoproteins are requisite for fundamental proteomics studies. The analysis of glycan structural alterations in glycoproteins is becoming increasingly important in terms of biomarkers, quality control of glycoprotein drugs, and the development of new drugs. However, usual approach such as proteoglycomics, glycoproteomics and glycomics which characterizes and/or identifies sugar chains, provides some structural information, but it does not provide any information of functionality of sugar chains. Therefore, in order to elucidate the function of glycans, functional glycomics which identifies the target glycoproteins and characterizes functional roles of sugar chains represents a promising approach. In this review, we show examples of functional glycomics technique using alpha 1,6 fucosyltransferase gene (Fut8) in order to identify the target glycoprotein(s). This approach is based on glycan profiling by CE/MS and LC/MS followed by proteomic approaches, including 2-DE/1-DE and lectin blot techniques and identification of functional changes of sugar chains.
Kondo
Akihiro
A
Department of Glycotherapeutics, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan. kondoa@glycot.med.osaka-u.ac.jp
Li
Wenzhe
W
Nakagawa
Takatoshi
T
Nakano
Miyako
M
Koyama
Nobuto
N
Wang
Xiangchun
X
Gu
Jianguo
J
Miyoshi
Eiji
E
Taniguchi
Naoyuki
N
D000818:Animals; D005647:Fucosyltransferases / Q000737:chemistry* / Q000235:genetics; D018390:Gene Targeting / Q000379:methods*; D023281:Genomics / Q000379:methods*; D006023:Glycoproteins / Q000302:isolation & purification; D006031:Glycosylation; D037102:Lectins / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D018345:Mice, Knockout; D040901:Proteomics / Q000379:methods; D034622:RNA Interference
D006023:Glycoproteins; D037102:Lectins; D005647:Fucosyltransferases; C020559:Glycoprotein 6-alpha-L-fucosyltransferase