Identification of a GH110 Subfamily of α1,3-Galactosidases: NOVEL ENZYMES FOR REMOVAL OF THE α3GAL XENOTRANSPLANTATION ANTIGEN

In search of α-galactosidases with improved kinetic properties for removal of the immunodominant α1,3-linked galactose residues of blood group B antigens, we recently identified a novel prokaryotic family of α-galactosidases (CAZy GH110) with highly restricted substrate specificity and neutral pH op...

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Published inThe Journal of biological chemistry Vol. 283; no. 13; pp. 8545 - 8554
Main Authors Liu, Qiyong P, Yuan, Huaiping, Bennett, Eric P, Levery, Steven B, Nudelman, Edward, Spence, Jean, Pietz, Greg, Saunders, Kristen, White, Thayer, Olsson, Martin L, Henrissat, Bernard, Sulzenbacher, Gerlind, Clausen, Henrik
Format Journal Article
LanguageEnglish
Published American Society for Biochemistry and Molecular Biology 28.03.2008
Subjects
Glycobiology and Extracellular Matrices
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Summary:In search of α-galactosidases with improved kinetic properties for removal of the immunodominant α1,3-linked galactose residues of blood group B antigens, we recently identified a novel prokaryotic family of α-galactosidases (CAZy GH110) with highly restricted substrate specificity and neutral pH optimum (Liu, Q. P., Sulzenbacher, G., Yuan, H., Bennett, E. P., Pietz, G., Saunders, K., Spence, J., Nudelman, E., Levery, S. B., White, T., Neveu, J. M., Lane, W. S., Bourne, Y., Olsson, M. L., Henrissat, B., and Clausen, H. (2007) Nat. Biotechnol. 25, 454-464). One member of this family from Bacteroides fragilis had exquisite substrate specificity for the branched blood group B structure Galα1-3(Fucα1-2)Gal, whereas linear oligosaccharides terminated by α1,3-linked galactose such as the immunodominant xenotransplantation epitope Galα1-3Galβ1-4GlcNAc did not serve as substrates. Here we demonstrate the existence of two distinct subfamilies of GH110 in B. fragilis and thetaiotaomicron strains. Members of one subfamily have exclusive specificity for the branched blood group B structures, whereas members of a newly identified subfamily represent linkage specific α1,3-galactosidases that act equally well on both branched blood group B and linear α1,3Gal structures. We determined by one-dimensional ¹H NMR spectroscopy that GH110 enzymes function with an inverting mechanism, which is in striking contrast to all other known α-galactosidases that use a retaining mechanism. The novel GH110 subfamily offers enzymes with highly improved performance in enzymatic removal of the immunodominant α3Gal xenotransplantation epitope.
Bibliography:The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EBI Data Bank with accession number(s) AM259273.
To whom correspondence should be addressed: ZymeQuest Inc., 100 Cummings Center, Suite 436H, Beverly, MA 01915-6122. Tel.: 978-232-8370; Fax: 978-232-8371; E-mail: pliu@zymequest.com.
This work was supported by ZymeQuest Inc. Work performed in M. L. O.'s laboratory was supported by the Swedish Research Council (project no. K2005-71X-14251), governmental ALF research grants to Lund University Hospital, and the Inga and John Hain Foundation for Medical Research and Region Skåne, Sweden. Work in H. C.'s laboratory was supported by the Danish Research Council and the Benzon Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The on-line version of this article (available at http://www.jbc.org) contains supplemental text, Figs. S1 and S2, Table S1, and additional references.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M709020200