Glycobiology, 2002, Vol. 12, No. 9 563-571
© 2002 Oxford University Press
Constitutively unmasked CD22 on B cells of ST6Gal I knockout mice: novel sialoside probe for murine CD22
2 Department of Molecular Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., MEM-L71, La Jolla, CA 92075, USA; 3 Shemyakin Institute of Bioorganic Chemistry, ul Miklukho-Maklaya 16/10, 117997 GSP-7, V-437 Moscow, Russian Federation; 4 Institute of Virology and Immunobiology, University of Würzburg, Versbacherstr. 7, 97078 Würzburg, Germany; and 5 HHMI Department of Cellular and Molecular Medicine, University of California San Diego, CMM-W Bldg., Room 333, 9500 Gilman Dr., 0625, La Jolla, CA 92073, USA
The interaction of CD22 with glycoprotein ligands bearing the Sia
2,6Gal-R sequence is believed to modulate its function as a regulator of B cell signaling. Although a commercial sialoside-polyacrylamide (PAA) probe, NeuAc- 2,6Gal-PAA, has facilitated studies on ligand binding by human CD22, murine CD22 binds instead with high affinity to NeuGc2,6Gal-R. A multivalent probe with this sequence was constructed to facilitate investigations of ligand binding in CD22 function using genetically defined murine models. The probe is based on the sialoside-PAA platform, which is then biotinylated for easy detection. A series of sialoside probes were constructed with two different length linker arms between the sialoside and the backbone and three different sialoside to PAA molar ratios. The NeuGc2,6Gal-PAA probe is specific for CD22: it binds to sialidase-treated B cells of wild-type mice but not B cells of CD22-null mice. Additionally, because the probe only binds to sialidase-treated wild-type cells, it confirms that CD22 is constitutively "masked" on most B cells from wild-type mice by binding to ligands in cis. In contrast, the probe bound equally well to native or sialidase-treated B cells from the immunocompromised ligand-deficient ST6Gal I knockout mice, demonstrating that CD22 is constitutively "unmasked" in these cells.
1 To whom correspondence should be addressed; E-mail: jpaulson{at}scripps.edu
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