Tags

Type your tag names separated by a space and hit enter

Characterization of sialyloligosaccharide binding by recombinant soluble and native cell-associated CD22. Evidence for a minimal structural recognition motif and the potential importance of multisite binding.
J Biol Chem. 1995 Mar 31; 270(13):7523-32.JB

Abstract

CD22, a B cell-specific receptor of the immunoglobulin superfamily, has been demonstrated to bind to oligosaccharides containing alpha 2-6-linked sialic acid (Sia) residues. Previously, we demonstrated that the minimal structure recognized by this lectin is the trisaccharide Sia alpha 2-6Gal beta 1-4GlcNAc, as found on N-linked, O-linked, or glycolipid structures (Powell, L., and Varki, A. (1994) J. Biol. Chem. 269, 10628-10636). Here we utilize a soluble immunoglobulin fusion construct (CD22Rg) to determine directly by equilibrium dialysis the stoichiometry (2:1) and dissociation constant (32 microM) for Neu5Ac alpha 2-6Gal beta 1-4Glc binding. Inhibition assays performed with over 30 different natural and synthetic sialylated and/or sulfated compounds are utilized to define in greater detail specific structural features involved in oligosaccharide-protein binding. Specifically, the critical features required for binding include the exocyclic hydroxylated side chain of the Sia residue and the alpha 2-6 linkage position to the underlying Gal unit. Surprisingly, alterations of the 2-, 3-, and 4-positions of the latter residue have limited effect on the binding. The nature of the residue to which the Gal is attached may affect binding. Bi(alpha 2-6)-sialylated biantennary oligosaccharides are capable of simultaneously interacting with both lectin sites present on the dimeric CD22Rg fusion construct, giving a marked improvement in binding over monosialylated compounds. Furthermore, data are presented indicating that full-length native CD22, expressed on the surface of Chinese hamster ovary cells, is structurally and functionally a multimeric protein, demonstrating a higher apparent affinity for multiply sialylated compounds over monosialylated compounds. These observations provide a mechanism for strong CD22-dependent cell adhesion despite the relatively low Kd for protein-sugar binding.

Authors+Show Affiliations

Department of Medicine, University of California at San Diego, La Jolla 92093, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

7706299

Citation

Powell, L D., et al. "Characterization of Sialyloligosaccharide Binding By Recombinant Soluble and Native Cell-associated CD22. Evidence for a Minimal Structural Recognition Motif and the Potential Importance of Multisite Binding." The Journal of Biological Chemistry, vol. 270, no. 13, 1995, pp. 7523-32.
Powell LD, Jain RK, Matta KL, et al. Characterization of sialyloligosaccharide binding by recombinant soluble and native cell-associated CD22. Evidence for a minimal structural recognition motif and the potential importance of multisite binding. J Biol Chem. 1995;270(13):7523-32.
Powell, L. D., Jain, R. K., Matta, K. L., Sabesan, S., & Varki, A. (1995). Characterization of sialyloligosaccharide binding by recombinant soluble and native cell-associated CD22. Evidence for a minimal structural recognition motif and the potential importance of multisite binding. The Journal of Biological Chemistry, 270(13), 7523-32.
Powell LD, et al. Characterization of Sialyloligosaccharide Binding By Recombinant Soluble and Native Cell-associated CD22. Evidence for a Minimal Structural Recognition Motif and the Potential Importance of Multisite Binding. J Biol Chem. 1995 Mar 31;270(13):7523-32. PubMed PMID: 7706299.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of sialyloligosaccharide binding by recombinant soluble and native cell-associated CD22. Evidence for a minimal structural recognition motif and the potential importance of multisite binding. AU - Powell,L D, AU - Jain,R K, AU - Matta,K L, AU - Sabesan,S, AU - Varki,A, PY - 1995/3/31/pubmed PY - 1995/3/31/medline PY - 1995/3/31/entrez SP - 7523 EP - 32 JF - The Journal of biological chemistry JO - J Biol Chem VL - 270 IS - 13 N2 - CD22, a B cell-specific receptor of the immunoglobulin superfamily, has been demonstrated to bind to oligosaccharides containing alpha 2-6-linked sialic acid (Sia) residues. Previously, we demonstrated that the minimal structure recognized by this lectin is the trisaccharide Sia alpha 2-6Gal beta 1-4GlcNAc, as found on N-linked, O-linked, or glycolipid structures (Powell, L., and Varki, A. (1994) J. Biol. Chem. 269, 10628-10636). Here we utilize a soluble immunoglobulin fusion construct (CD22Rg) to determine directly by equilibrium dialysis the stoichiometry (2:1) and dissociation constant (32 microM) for Neu5Ac alpha 2-6Gal beta 1-4Glc binding. Inhibition assays performed with over 30 different natural and synthetic sialylated and/or sulfated compounds are utilized to define in greater detail specific structural features involved in oligosaccharide-protein binding. Specifically, the critical features required for binding include the exocyclic hydroxylated side chain of the Sia residue and the alpha 2-6 linkage position to the underlying Gal unit. Surprisingly, alterations of the 2-, 3-, and 4-positions of the latter residue have limited effect on the binding. The nature of the residue to which the Gal is attached may affect binding. Bi(alpha 2-6)-sialylated biantennary oligosaccharides are capable of simultaneously interacting with both lectin sites present on the dimeric CD22Rg fusion construct, giving a marked improvement in binding over monosialylated compounds. Furthermore, data are presented indicating that full-length native CD22, expressed on the surface of Chinese hamster ovary cells, is structurally and functionally a multimeric protein, demonstrating a higher apparent affinity for multiply sialylated compounds over monosialylated compounds. These observations provide a mechanism for strong CD22-dependent cell adhesion despite the relatively low Kd for protein-sugar binding. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/7706299/Characterization_of_sialyloligosaccharide_binding_by_recombinant_soluble_and_native_cell_associated_CD22__Evidence_for_a_minimal_structural_recognition_motif_and_the_potential_importance_of_multisite_binding_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(18)71793-6 DB - PRIME DP - Unbound Medicine ER -