Comparisons of Caenorhabditis Fucosyltransferase Mutants Reveal a Multiplicity of Isomeric N-Glycan Structures

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• 作者：Shi Yan ; Chunsheng Jin ; Iain B. H. Wilson ; Katharina Paschinger
• 刊名：Journal of Proteome Research
• 出版年：2015
• 出版时间：December 4, 2015
• 年：2015
• 卷：14
• 期：12
• 页码：5291-5305
• 全文大小：1263K
• ISSN：1535-3907

文摘

Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core 伪1,6-fucose as well as the presence of fucose on the bisecting galactose. In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. These data not only show the high variability of the N-glycomic capacity of a 鈥渟imple鈥?nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes.