In this approach, we discuss the glycome of C.elegans in relation to N-glycan profiling obtained from tissue extraction, with glycans released by enzymatic and chemical methods, and components structures detailed by mass spectrometry.

Protein extract from mixed worm population was submitted to Pngase F, A and hydrazinolysis for N-glycans releasing. They were permethylated and analyzed by mass spectrometry. Each method was complementary, and the Pngase A was 3-5 times more efficient to remove N-glycan from C.elegans than Pngase F using standard isotopic dilution approach with mass shifts and relative quantification provided by glycan deuteromethyl analog. Only hydrazinolysis was able to release several highly fucosylated glycans. These glycans represented a series of glycoforms which included high mannose type (Hex_5-9HexNAc₂), as well as several truncated and extended fucosylated glycans (Hex_3-7HexNAc₂Fuc_0-4). N-complex glycan type found in mammals were not found in C.elegans. To address if the truncated forms were products of action of beta-N-acetyl-glucosaminidase activity, we introduced an exogenous glycoprotein, asialo-fetuin, to protein extract. N-glycans from asialo-fetuin were removed by Pnase F in presence and absence of worm extract and released N-glycans were labeled with deutero/non-deuterated-methyl iodide for mass spectrometry quantification. The ratio of complex try-antennary glycan from asialo-fetuin was recovery quantitatively which demonstrated that none appreciable degradation occurred. Previous investigations of the N-glycans in C.elegans, including our own, have frequently employed global N-glycan release, but not addressed to entities that glycans are coupled, the glycoproteins. To step next level, we have been stained C.elegans with FITC-labeled lectins and showed that KM+, jacalin and lentil lectin stained differentially parts of C.elegans body and the isolation of subpopulation of glycoproteins using affinity capture by these lectins is underway. FAPESP, CAPES and FAEPA.