Sulfation and sialylation requirements for a glycoform of CD34, a major endothelial ligand for L-selectin in porcine peripheral lymph nodes

K. Shailubhai, Philip Streeter, C. E. Smith, G. S. Jacob

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Leukocyte recruitment from blood into peripheral lymph nodes is controlled in part by a specific interaction of lymphocyte-associated L-selectin with endothelial cell receptors known as peripheral addressins. In murine lymph nodes, two peripheral addressins have been identified, Gly-CAM-1, a 50 kDa molecule that also appears as a secreted form in plasma, and CD34, a 90 kDa membrane-associated sialomucin. A predominant 105 kDa CD34 mucin-like protein has also been identified in human tonsil as peripheral addressin. We have identified a 120 kDa sialomucin as the predominant peripheral addressin in porcine lymph nodes. Validation of the 120 kDa porcine molecule as a peripheral addressin was based on its ability to bind MECA-79, a monoclonal antibody previously used to isolate peripheral addressins from mouse and human tissues, and to bind an L-selectin-Fc chimera (LS-Fc). The binding with LS-Fc was abolished in the presence of fucoidin, a sulfated polysaccharide known to inhibit L-selectin-receptor interactions. To address the possibility that the 120 kDa ligand may contain common recognition determinants for MECA-79 and L-selectin, the requirements for sialylation and sulfation were compared. Whereas desialylation of 120 kDa ligand drastically-reduced its binding to LS-Fc, this treatment appeared to enhance the binding of 120 kDa ligand to MECA-79. In contrast, the binding of both MECA-79 and LS-Fc to 120 kDa ligand was drastically reduced when de novo sulfation of this ligand was reduced by including chlorate, a metabolic inhibitor of sulfation, in the culture media. N-Terminal amino acid sequences of the porcine 120 kDa protein revealed homology with human CD34. Taken together, these findings suggest that the porcine 120 kDa peripheral addressin is an L-selectin-binding glycoform of CD34.

Original languageEnglish (US)
Pages (from-to)305-314
Number of pages10
JournalGlycobiology
Volume7
Issue number2
DOIs
StatePublished - Mar 1997
Externally publishedYes

Fingerprint

L-Selectin
Swine
Lymph Nodes
Ligands
Sialomucins
Chlorates
Computer peripheral equipment
Molecules
Lymphocytes
Palatine Tonsil
Endothelial cells
Mucins
Computer aided manufacturing
Polysaccharides
Culture Media
Amino Acid Sequence
Proteins
Leukocytes
Blood
Endothelial Cells

Keywords

  • CD34
  • Inflammation
  • L-Selectin
  • Sialylation
  • Sulfation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sulfation and sialylation requirements for a glycoform of CD34, a major endothelial ligand for L-selectin in porcine peripheral lymph nodes. / Shailubhai, K.; Streeter, Philip; Smith, C. E.; Jacob, G. S.

In: Glycobiology, Vol. 7, No. 2, 03.1997, p. 305-314.

Research output: Contribution to journalArticle

Shailubhai, K. ; Streeter, Philip ; Smith, C. E. ; Jacob, G. S. / Sulfation and sialylation requirements for a glycoform of CD34, a major endothelial ligand for L-selectin in porcine peripheral lymph nodes. In: Glycobiology. 1997 ; Vol. 7, No. 2. pp. 305-314.
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