Posttranslational processing of proopiomelanocortin family molecules in sea lamprey based on mass spectrometric and chemical analyses.
In gnathostomes, adrenocorticotropic hormone (ACTH), melanophore-stimulating hormones (MSHs), and beta-endorphin (beta-END) are derived from a common precursor, proopiomelanocortin. In sea lamprey, ACTH and two forms of MSHs are contained in independent precursors, proopiocortin (POC), and proopiomelanotropin (POM), respectively, together with a distinct beta-END. Here, we characterized products from POC and POM. An analysis of previously purified ACTH by mass spectrometry (MS) detected four peptides with a molecular weight of 6469.4, 6549.6, 6556.6, or 6636.1. The sequence analysis of an ACTH preparation following enzymatic and chemical cleavage revealed the presence of ACTH(1-59) and ACTH(1-60) corresponding to a molecular weight of 6469.4 and 6556.6, respectively, and of ACTH(1-59) and ACTH(1-60) modified at Ser(35) by a group having a mass of 80, giving the molecular weight 6549.6 and 6636.1, respectively. The modification could be due to phosphorylation based on the increase in molecular weight of 80. Analyses of frozen pituitary slices with MALDI-TOF MS detected several mass numbers corresponding to POC-derived peptides such as ACTH(1-60), modified ACTH(1-60), and (POC)beta-END, and those corresponding to POM-derived peptides such as MSH-A, MSH-B, and the C-terminal fragment of (POM)beta-END lacking a Met-enkephalin segment. The present results together with previous characterizations show that in sea lamprey pituitary the major products derived from POC in the PD by posttranslational processing are ACTH and beta-END as in gnathostomes. The posttranslational processing of POM in the PI is similar to that in gnathostomes in the sense of the occurrence of MSH, however, it differs in that beta-END is further cleaved, thus generating Met-enkephalin.
Takahashi
Akiyoshi
A
Laboratory of Molecular Endocrinology, School of Fisheries Sciences, Kitasato University, Sanriku, Ofunato, Iwate 022-0101, Japan. akiyoshi@kitasato-u.ac.jp
Yasuda
Akikazu
A
Sower
Stacia A
SA
Kawauchi
Hiroshi
H
D000324:Adrenocorticotropic Hormone / Q000737:chemistry; D000818:Animals; D005629:Frozen Sections; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D048428:Petromyzon / Q000378:metabolism*; D010902:Pituitary Gland / Q000737:chemistry; D011333:Pro-Opiomelanocortin / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000521:alpha-MSH / Q000737:chemistry; D019824:beta-MSH / Q000737:chemistry
D010446:Peptide Fragments; D019824:beta-MSH; D000521:alpha-MSH; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone