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J . Org. Chem. 2001, 66, 5248-5251
Acid -Ca ta lyzed Hyd r olysis a n d
La cton iza tion of r2,8-Lin k ed Oligosia lic
Acid s
†
‡
‡
Yi-Ping Yu, Mou-Chi Cheng, Heng-Ru Lin,
Chun-Hung Lin,† and Shih-Hsiung Wu*
,‡,§
,†,‡,§
Graduate Institute of Life Sciences, National Defense
Medical Center; Institute of Biological Chemistry, Academia
Sinica; and Graduate Institute of Biochemical Sciences,
National Taiwan University, Taipei, Taiwan
Received February 19, 2001
Polysialic acids (PSA) exist mainly on the surface of
mammalian cells and certain bacteria and are involved
1
in many important biological roles. For example, the
R2,8-PSA chain is expressed in neural cell adhesion
molecules (N-CAM) in a developmentally regulated man-
ner. The homopolymers of N-acetylneuraminic acids have
been implicated in altering the shape and movement of
cells because PSA can attenuate adhesion forces and
2
modulate overall cell surface interaction. There are two
distinct features of PSA in relation to its critical physi-
ological roles: intramolecular lactonization and the labil-
ity of glycosidic linkages. The former influences the
charge distribution and tertiary structure of PSA, and
the latter varies the size of PSA-containing biopolymers.
Of all the glycosidic linkages found in oligosaccharides,
3
those of polysialic acids are remarkbly labile. R-2,8-
Polysialic acid has been noted for its degradation in
prolonged freezer storage4 and in the preparation of
5
N-CAM for SDS gel anlaysis. Acid-catalyzed cleavage
of the R2,8-linkage in oligo- and polysialic acids (OSA/
PSA) was reported to occur preferentially at the linkage
between two internal sialic acid residues. The cleavage
of the internal R2,8-linkages in PSA is much easier as a
*
Fax: +886-2-2653-9142.
National Defense Medical Center.
Academia Sinica.
†
F igu r e 1. Structures of R2,8-linked sialic acid tetramer 1 and
partially and fully lactonized tetramer 5.
‡
§
National Taiwan University.
(
1) (a) Troy, F. A., II. Glycobiology 1992, 2, 5-23. (b) Rutishauser,
result of the intramolecular catalysis of general acid from
U.; Acheson, A.; Hall, A. K.; Mann, D. M.; Sunshine, J . Science 1988,
a
the adjacent carboxyl groups that have high pK ’s as
2
40, 53-57. (c) Kitajima, K.; Inoue, S.; Inoue, Y.; Troy, F. A., II. J .
4
proton donors. Intramolecular lactonization of OSA/PSA
(Figure 1) was also easily catalyzed by acid and occurred
preferentially at the linkage between two internal sialic
acid residues. The lactonization process displays three
Biol. Chem. 1988, 263, 18269-18276. (d) Zuber, C.; Lackie, P. M.;
Caterall, W. A.; Roth, J . J . Biol. Chem. 1992, 267, 9965-9971. (e)
Inoue, S.; Iwasaki, M. Biochem. Biophys. Res. Commun. 1978, 83,
1
018-1023. (f) Robbin, J . B.; McCracken, G. H., J r.; Gotschlich, E. C.;
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220. (g) Schiffer, M. S.; Oliverira, E.; Glode, M. P.; McCracken, G.
1
discrete stages from limited lactonization to full lacton-
H., J r.; Sarff, L. M.; Robbin, J . B. Pediatr. Res. 1976, 10, 82-87. (h)
Baumann, H.; Brisson, J . R.; Michon, F.; Pon, R.; J enning, H. J .
Biochemistry 1993, 32, 4007-4013. (i) Hayrinen, J .; J enning, H. J .;
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ization.6 Therefore, lactonization and hydrolysis are
competitive under acidic conditions. We have previously
shown that lactonized species of OSA/PSA, generated
from the hydrolysis of colominic acid under acidic condi-
tions, severely interfere with their separation in capillary
electrophoresis, but their existence could be detected by
(2) (a) Rutishhauser, U.; Landmesser, L. Trends Neurosci. 1996, 19,
4
6
6
22-427. (b) Kiss, J . Z.; Rougon, G. Curr. Opin. Neurobiol. 1997, 7,
40-646. (c) Rutishhauser, U. Curr. Opin. Cell Biol. 1996, 8, 679-
84. (d) Shen, H.; Watanabe, M.; Tomasiewicz, H.; Rutishhauser, U.;
7
mass spectrometry or high performance anion-exchange
Magnuson, T.; Glass, J . D. J . Neurosci. 1997, 17, 5221-5229.
3) Schauer, R. In Sialic Acids: Chemistry, Metabolism and Func-
tion, Cell Biology Monographs; Springer-Verlag: New York, 1982; Vol.
(
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Chem. 2000, 112, 788-792; Angew. Chem., Int. Ed. 2000, 39, 772-
776.
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Biochem. 1998, 260, 154-159. (b) Cheng, M. C.; Wang, K. T.; Inoue,
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1
0.
(
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2
(
Mailhammer, R.; Rutishauer, U.; Cunningham, B. A.; Edelman, G. M.
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1
0.1021/jo015580c CCC: $20.00 © 2001 American Chemical Society
Published on Web 06/29/2001