Isolation and characterization of three forms of joining peptide from adult human pituitaries: lack of adrenal androgen-stimulating activity.
Three structural variants of the joining peptide (JP) fragment of POMC have been purified from human pituitaries. Ion exchange and reverse phase tissue extraction procedures were combined with reverse phase HPLC to achieve complete purification of each form of JP. Fragments resulting from tryptic hydrolysis of each form were characterized by amino acid analysis and fast atom bombardment mass spectrometry. The predominant form of human JP, accounting for about 50% of the total purified, was found to be conjugated to glutathione through the lone cysteine residue at position 9. The other two variants were identified as human JP with a free cysteine residue and human JP dimer and accounted for 35% and 15%, respectively, of the total purified. Recently, human JP-(1-18) has been suggested as having adrenal androgen-stimulating activity. None of the three JP variants or their respective 1-20 amino-terminal fragments resulting from tryptic hydrolysis showed any ability to promote the secretion of dehydroepiandrosterone sulfate by cultured human fetal adrenal cells. Similarly, no potentiation of the stimulatory effects of ACTH-(1-39) was observed. The three variants of human JP as well as JP purified from rat, porcine, and bovine pituitaries were tested for their ability to stimulate androgenic steroids from dispersed fetal rabbit adrenal cells. None showed any significant biological activity either in stimulating steroid secretion or in potentiating the action of ACTH-(1-39).
Robinson
P
P
Endocrine Laboratory, Royal Victoria Hospital, Montreal, Quebec, Canada.
Bateman
A
A
Mulay
S
S
Spencer
S J
SJ
Jaffe
R B
RB
Solomon
S
S
Bennett
H P
HP
D000311:Adrenal Glands / Q000187:drug effects* / Q000196:embryology / Q000378:metabolism; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives* / Q000378:metabolism; D019314:Dehydroepiandrosterone Sulfate; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D010902:Pituitary Gland / Q000737:chemistry*; D011333:Pro-Opiomelanocortin / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013552:Swine; D014357:Trypsin
D046911:Macromolecular Substances; D010446:Peptide Fragments; D003687:Dehydroepiandrosterone; D019314:Dehydroepiandrosterone Sulfate; D011333:Pro-Opiomelanocortin; D014357:Trypsin