Determination of the glycoforms of human chorionic gonadotropin b-core fragment by matrix-assisted laser desoption/ionisation time of flight mass spectrometry
Jacoby, Eli S. and Kicman, Andrew T. and Laidler, Paul and Iles, Ray K. (2000) Determination of the glycoforms of human chorionic gonadotropin b-core fragment by matrix-assisted laser desoption/ionisation time of flight mass spectrometry. Clinical Chemistry, 46 (11). pp. 1796-1803. ISSN 0009-9147
Background: Metabolism of human chorionic gonadotropin (hCG) in the serum and kidney yields the terminal urinary product hCG ß-core fragment (hCGßcf), comprising two disulfide-linked peptides (ß6-ß40 and ß55-ß92) of which one (ß6-ß40) retains truncated N-linked sugars. Hyperglycosylated hCGßcf may indicate choriocarcinoma or Down syndrome, but the glycosylation profile of hCGßcf has not been thoroughly evaluated.
Methods: hCGßcf, purified from pregnancy urine, was reduced by "on-target" dithiothreitol (DTT) reduction and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The mass ([M+H]+) of the primary sequence of the glycosylated peptide ß6-ß40 was subtracted from the m/z values of the discrete peaks observed to give the masses of the carbohydrate moieties. Carbohydrate structure was predicted by sequentially subtracting the masses of the monosaccharide residues corresponding to N-linked carbohydrates of the hCG ß-subunit reported in the literature.
Results: Mass spectra of hCGßcf revealed a broad triple peak at m/z 8700–11300. After reduction, the triple peak was replaced by a discrete set of peaks between m/z 4156 and 6354. A peak at m/z 4156.8 corresponded to the nonglycosylated peptide (ß55-ß92). The remaining nine peaks indicated that urinary hCGßcf comprises a set of glycoforms smaller and larger than the trimannosyl core.
Conclusions: hCGßcf comprises a wider set of glycoforms than reported previously. Peaks of highest mass indicate evidence of hyperglycosylated carbohydrate moieties. The data support previous reports that hCGßcf oligosaccharides lack sialic acid and galactose residues. No indication was found of a ß6-ß40 peptide that was entirely devoid of carbohydrate.
|Research Areas:||A. > School of Science and Technology > Natural Sciences|
|Depositing User:||Repository team|
|Date Deposited:||21 May 2009 08:21|
|Last Modified:||15 Feb 2016 15:06|
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