Identification of a tyrosine residue in ovine placental lactogen as essential for its binding to receptors.
Nitration of ovine placental lactogen (oPL) with a 10-fold molar excess of tetranitromethane over protein content resulted in the modification of 0.8 tyrosine residue. No conformational changes were observed by either fourth-derivative spectral analysis or circular dichroism. Nitration significantly decreased the binding capacity of the hormone to lactogenic and somatogenic rat liver receptors. This binding capacity was not restored by reduction of the nitro groups, thus indicating that the decrease was not due to the difference in pK between tyrosine and nitrotyrosine. The nitrotyrosine-containing peptide was isolated from a tryptic digest by HPLC and its modification extent was of 67%, which is consistent with the decrease in binding capacities (65% and 70%). Its amino acid sequence was determined and the modified tyrosine residue was identified as Tyr-46. These results provide the first evidence of the involvement of a tyrosine residue in the binding of oPL to both lactogenic and somatogenic receptors. This tyrosine appears to be a shared binding epitope between oPL and the prolactins.
Cymes
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Instituto de Química y Fisicoquímica Biólogicas, Buenos Aires, Argentina.
Wolfenstein-Todel
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D005260:Female; D013006:Growth Hormone / Q000037:antagonists & inhibitors / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010928:Placental Lactogen / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D051381:Rats; D018000:Receptors, Peptide / Q000378:metabolism*; D011986:Receptors, Somatotropin / Q000378:metabolism*; D012756:Sheep; D013053:Spectrophotometry; D013774:Tetranitromethane / Q000737:chemistry; D014357:Trypsin / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
D010446:Peptide Fragments; D018000:Receptors, Peptide; D011986:Receptors, Somatotropin; C028202:placental lactogen receptor; D014443:Tyrosine; D013006:Growth Hormone; D010928:Placental Lactogen; D014357:Trypsin; D013774:Tetranitromethane