The molecular characterisation of chicken pituitary N-terminal pro-opiomelanocortin (POMC).
Monoclonal antibodies (Mabs) specifically recognizing the chicken pituitary corticotropes were used to isolate a population of closely related peptides from crude chicken pituitary extracts. A homogeneous N-terminal sequence homologous to the extreme N-terminus of mammalian and amphibian pro-opiomelanocortin (POMC) was revealed. Further physicochemical analysis proved the existence of a series of C-terminally truncated peptides including 3 major molecular species corresponding to Ser1-Gly64, Ser1-Arg73 and Ser1-Gly105 respectively. The two latter molecules were shown to be N-glycosylated at position Asn67, with mass spectrometric data indicating a carbohydrate structure of the oligomannose 5 type, in addition to two more complex structures. No evidence was found in favour of O-glycosylation on Ser47. Degenerated PCR primers were deduced from the above protein sequence and from the known chicken adrenocorticotropic hormone (ACTH) sequence. The nucleotide sequence obtained by reversed transcription PCR (RT-PCR) completely confirmed the new amino acid sequence data including pro-gamma-MSH, the joining peptide and ACTH.
Berghman
L R
LR
Laboratory of Neuroendocrinology and Immunological Biotechnology, Zoological Institute, Leuven, Belgium. Luc.Berghman@bio.KULeuven.ac.be
Devreese
B
B
Verhaert
P
P
Gerets
H
H
Arckens
L
L
Vanden Broeck
J
J
Van Beeumen
J
J
Vaudry
H
H
Vandesande
F
F
D000324:Adrenocorticotropic Hormone / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002645:Chickens / Q000235:genetics*; D006031:Glycosylation; D007150:Immunohistochemistry; D008358:Mannose / Q000032:analysis; D013058:Mass Spectrometry; D009074:Melanocyte-Stimulating Hormones / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D010902:Pituitary Gland / Q000737:chemistry*; D011333:Pro-Opiomelanocortin / Q000737:chemistry / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017385:Sequence Homology
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone; D009074:Melanocyte-Stimulating Hormones; D008358:Mannose