Thyroid hormone regulation of peptidylglycine alpha-amidating monooxygenase expression in anterior pituitary gland.
Peptidylglycine alpha-amidating monooxygenase (PAM; EC 1.14.17.3) is a copper-, molecular oxygen-, and ascorbate-dependent enzyme which catalyzes the COOH-terminal amidation of bioactive peptides. Expression of PAM in the adult male rat anterior pituitary was evaluated after experimental manipulation of thyroid status. Levels of PAM mRNA increased 4- to 7-fold in animals made hypothyroid by treatment with 6-n-propyl-2-thiouracil or thyroidectomy and were not diminished below control levels in animals made hyperthyroid by treatment with T4. Treatment of thyroidectomized animals with T4 prevented the increase in PAM mRNA levels; similar doses of T4 returned serum TSH and anterior pituitary PAM mRNA to euthyroid values. Based on Northern blot analysis and amplification of fragments derived from rat PAM-1 by reverse transcription and the polymerase chain reaction, thyroid status did not affect the distribution of PAM mRNA among its various alternatively spliced forms. The specific activity of PAM in the anterior pituitary was increased slightly in both the soluble and particulate fractions from chemically hypothyroid rats; the majority of the PAM activity in the rat anterior pituitary was soluble, and increased secretion of enzyme may account for the lesser effect of chemical thyroidectomy on specific activity compared to mRNA levels. Western blot analysis demonstrated a 104-kDa PAM protein in particulate fractions prepared from control, PTU-treated, and T4-treated animals. The soluble fraction contained major PAM proteins of 95 and 75 kDa, and PTU treatment brought about an increase in the prevalence of the 75-kDa form of PAM protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Ouafik
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Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
May
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Saffen
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D000818:Animals; D015153:Blotting, Western; D005786:Gene Expression Regulation / Q000187:drug effects; D007037:Hypothyroidism / Q000209:etiology / Q000378:metabolism; D008297:Male; D006899:Mixed Function Oxygenases / Q000235:genetics*; D009097:Multienzyme Complexes; D009693:Nucleic Acid Hybridization; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011441:Propylthiouracil; D012326:RNA Splicing; D012333:RNA, Messenger / Q000096:biosynthesis; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013963:Thyroid Hormones / Q000494:pharmacology / Q000502:physiology*; D013965:Thyroidectomy; D013972:Thyrotropin / Q000097:blood; D013974:Thyroxine / Q000494:pharmacology
D009097:Multienzyme Complexes; D012333:RNA, Messenger; D013963:Thyroid Hormones; D011441:Propylthiouracil; D013972:Thyrotropin; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D012194:RNA-Directed DNA Polymerase; D013974:Thyroxine