Heparin binding lectin of human placenta as a tool for histochemical ligand localization and isolation.
Biotinylated heparin has been used to detect the presence of specific binding sites in sections of human placenta, which has prompted demonstration of expression of lectin activity for this proteoglycan. Purification of this lectin from full-term placenta facilitates the synthesis of its biotinylated derivative, using biotin-amidocaproyl hydrazide, without affecting its activity. It also enables immunization to obtain antibodies. The labeled lectin is shown to bind specifically to nuclear and cytoplasmic locations in various cell types of human placenta, nuclear expression of lectin binding sites being more pronounced at the full-term stage than after 8 weeks of development. The structurally related histone H2B exhibits obvious differences in its binding pattern. The presence of ligands accessible to the lectin whose binding activity can be inhibited by addition of an excess of heparin correlates in most instances with the level of lectin expression detected immunohistochemically. Biochemical information on the nature of the glycohistochemically inferred lectin-specific ligand(s) is obtained by affinity chromatography on resin-immobilized lectin. It leads to isolation of a proteoglycan with similar electrophoretic mobility in agarose-polyacrylamide gel electrophoresis relative to the independently purified heparan sulfate-containing fibronectin binding proteoglycan from human placenta. Both fractions inhibit binding of heparin to the lectin and contain immunologically detected co-purified lectin, emphasizing their ligand properties. Application of labeled tissue lectins in conjunction with lectin-specific antibodies is proposed to obtain valuable insights into the expression of the receptor as well as the ligand part of protein-carbohydrate recognition.
Max-Planck-Institut für Experimentelle Medizin, Abteilung Chemie, Göttingen, Federal Republic of Germany.
H J
HJ
Gabius
B
B
Kohnke-Godt
M
M
Leichsenring
A
A
Bardosi
D002846:Chromatography, Affinity; D004586:Electrophoresis / Q000379:methods; D005260:Female; D006493:Heparin / Q000378:metabolism*; D006651:Histocytochemistry; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D008024:Ligands; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D011485:Protein Binding; D011509:Proteoglycans / Q000378:metabolism; D011975:Receptors, Mitogen / Q000032:analysis*
D037102:Lectins; D008024:Ligands; D011509:Proteoglycans; D011975:Receptors, Mitogen; D006493:Heparin