Phosphorylated forms of adrenocorticotropin and corticotropin-like intermediary lobe peptide in human tumors.
Many peptides contribute to the heterogeneity of immunoreactive adrenocorticotropin (ACTH) in man. The use of a radioimmunoassay (RIA) specifically directed against the C-terminal end of ACTH allowed us to study precisely the following four peptides: ACTH itself, corticotropin-like intermediary lobe peptide (CLIP) or ACTH (18-39) and their phosphorylated forms on Ser31. We have set up a high-performance liquid chromatography system that separates these four molecules in a single run, to establish their relative distributions in tumors responsible for Cushing's disease or for the ectopic ACTH syndrome, and to evaluate the possible interference of phospho-Ser31 on various RIA or immunoradiometric assay (IRMA) recognition systems for ACTH. In this system, alkaline phosphatase treatment shifted the retention time of the phosphorylated peptides to that of their non-phosphorylated counterparts. In three tumors responsible for the ectopic ACTH syndrome, CLIP peptides were predominant in two and phosphorylated molecules represented between 22% and 50% of immunoreactive materials. In five pituitary tumors responsible for Cushing's disease, ACTH peptides were predominant and the phosphorylated molecules varied between 35% and 75% in four of them. In the same tumor the ratios of phosphorylated to non-phosphorylated CLIP or ACTH were identical. The presence of phospho-Ser31 did not affect the recognition ability of two mid-ACTH and two C-terminal ACTH RIAs, nor of the ACTH IRMA (Allegro, Nichols).
Massias
J F
JF
INSERM CJF 92-08, Institut Cochin de Génétique Moléculaire, Faculté Cochin-Port Royal, Université René Descartes, Paris, France.
Hardouin
S
S
Vieau
D
D
Lenne
F
F
Bertagna
X
X
D000236:Adenoma / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D000310:Adrenal Gland Neoplasms / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D000324:Adrenocorticotropic Hormone / Q000032:analysis* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D053497:Corticotropin-Like Intermediate Lobe Peptide; D006801:Humans; D015592:Immunoradiometric Assay; D010446:Peptide Fragments / Q000032:analysis* / Q000378:metabolism*; D010673:Pheochromocytoma / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation; D010911:Pituitary Neoplasms / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D011863:Radioimmunoassay
D010446:Peptide Fragments; D053497:Corticotropin-Like Intermediate Lobe Peptide; D000324:Adrenocorticotropic Hormone