cAmp regulation of arylalkylamine N-acetyltransferase (AANAT, EC 2.3.1.87): a new cell line (1E7) provides evidence of intracellular AANAT activation.
Arylalkylamine N-acetyltransferase (serotonin N-acetyltransferase, AANAT, EC ) is the penultimate enzyme in melatonin synthesis. As described here, a cell line (1E7) expressing human AANAT (hAANAT) has been developed to study the human enzyme. 1E7 hAANAT is detectable in immunoblots as a 23-kDa band and is immunocytochemically visualized in the cytoplasm. The specific concentration of hAANAT in homogenates is comparable to that of the night rat pineal gland. Kinetics of AANAT extracted from 1E7 cells are the same as those of bacterially expressed hAANAT; both preparations of hAANAT are equally sensitive to the inhibitor CoA-S-N-acetyltryptamine. Studies of cAMP regulation indicate that treatment with forskolin, dibutyryl cAMP, isobutylmethylxanthine, or isoproterenol activate cellular hAANAT within intact 1E7 cells approximately 8-fold without markedly increasing the abundance of AANAT protein or the activity of AANAT in broken cell preparations; and, that forskolin, isobutylmethylxanthine and isoproterenol elevate cyclic AMP production. These observations extend our understanding of cAMP regulation of AANAT activity, because it is currently thought that this only involves changes in the steady-state levels of AANAT protein. This previously unrecognized switching mechanism could function physiologically to control melatonin production without changing AANAT protein levels.
Coon
S L
SL
Section on Neuroendocrinology, Laboratory of Developmental Neurobiology, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA.
Weller
J L
JL
Korf
H W
HW
Namboodiri
M A
MA
Rollag
M
M
Klein
D C
DC
D000818:Animals; D047068:Arylalkylamine N-Acetyltransferase; D001191:Arylamine N-Acetyltransferase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019556:COS Cells; D002457:Cell Extracts / Q000032:analysis; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology*; D003593:Cytoplasm / Q000201:enzymology; D003624:Darkness; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008550:Melatonin / Q000378:metabolism; D010870:Pineal Gland / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014363:Tryptamines / Q000494:pharmacology
D002457:Cell Extracts; D014363:Tryptamines; C043464:N-acetyltryptamine; D005576:Colforsin; D000242:Cyclic AMP; D001191:Arylamine N-Acetyltransferase; C478879:AANAT protein, human; D047068:Arylalkylamine N-Acetyltransferase; D008550:Melatonin