Change in the accessibility of an epitope of the human granulocyte-colony stimulating factor after binding to receptors.
In a previous work we demonstrated that monoclonal antibody (mAb) 8C2 recognized a human granulocyte-colony stimulating factor (hG-CSF) region left unmasked after binding to placenta receptors, whereas mAb 6E3 defined a receptor-buried epitope. Herein we examined the role of these antigenic regions on the proliferative response induced by hG-CSF on a myeloid leukaemia cell line. Both mAbs significantly inhibited the hG-CSF-induced cell growing, although epitope 8C2 but not 6E3 remained exposed in hG-CSF:cell receptor complexes. When cytokine:receptor complexes already formed at 4 degrees C were incubated 1 h at 37 degrees C under conditions preventing the internalization, a significant reduction in the amount of accessible 8C2 epitopes was evident. However, this effect was not observed when mAb 8C2:hG-CSF complexes previously bound to cells were incubated at 37 degrees C. Thus, results suggest that a receptor oligomerization process could account for the temperature-induced epitope 8C2 masking. The identification of epitope 8C2 accomplished by synthesis of overlapping octapeptides, revealed that it is formed by sequences 39-52 and 155-164, both in close proximity in the three-dimensional structure of the hG-CSF molecule. Since part of this region has been proposed as a second binding site to receptors, we infer that the change of epitope 8C2 accessibility could be the result of either receptor aggregation or epitope binding to another receptor. In addition, our data support the hypothesis that a ligand-induced receptor oligomerization is required for transduction of cytokine signals.
Marino
Verónica J
VJ
Facultad de Farmacia y Bioquímica, Instituto de Química y Fisicoquimica Biológicas (UBA-CONICET), Junin 956, 1113 Buenos Aires, Argentina.
Sterin-Prync
Aída E
AE
Roguin
Leonor P
LP
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D002455:Cell Division; D045744:Cell Line, Tumor; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D016188:Receptors, Granulocyte Colony-Stimulating Factor / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D013696:Temperature
D000911:Antibodies, Monoclonal; D000939:Epitopes; D016188:Receptors, Granulocyte Colony-Stimulating Factor; D016179:Granulocyte Colony-Stimulating Factor