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Journal of Lipid Research, Vol 30, 1781-1787, Copyright © 1989 by Lipid Research, Inc.
Y Ito, JL Breslow and BT Chait
Apolipoprotein C-III (apo C-III) is a 79 amino acid glycoprotein. The sugar
moiety of apoC-III is attached to amino acid residue 74 and is thought to
consist of 1 mole of galactose, 1 mole of N-acetyl- galactosamine, and
either 0, 1, or 2 moles of sialic acid. This results in three isoproteins
called C-III0, C-III1, and C-III2 designated by the number of sialic acid
residues. It has been assumed, although not experimentally tested, that
apoC-III0 lacks sialic acid residues but possesses the
D-galactosyl-(1-3)-N-acetyl-D-galactosamine sugar backbone. To verify the
structure of the three apoC-III isoproteins, we applied the method of 252Cf
plasma desorption mass spectrometry to measure the exact molecular weight
(Mr) of each of the isoproteins. Our data confirmed the proposed structure
of apoC-III1 and apoC-III2. However, the difference in mass between
apoC-III1 (9420.6, 9420.0, 9422.2 daltons) and apoC-III0 (8763.9, 8764.9,
8765.5 daltons, respectively, in three subjects) suggests that the latter
is missing not just sialic acid but the entire sugar moiety. This finding
may have important implications for the metabolism of apoC-III. The
accuracy and reproducibility of Mr measurements described in this paper
suggest that this technique holds promise for the detection of
apolipoprotein amino acid substitutions or modifications undetected by
conventional techniques such as isoelectric focusing.
ARTICLES
Apolipoprotein C-III0 lacks carbohydrate residues: use of mass spectrometry to study apolipoprotein structure
Laboratory of Biochemical Genetics and Metabolism, Rockefeller University, New York, NY 10021.
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