Characterization of the peptide acetyltransferase activity in bovine and rat intermediate pituitaries responsible for the acetylation of beta-endorphin and alpha-melanotropin.
Year: 1982
Type: article
Abstract: The peptide acetyltransferase activity responsible for the NH2-terminal acetylation of P-endorphin, adrenocorticotropic hormone (ACTH)(l-l3)NHZ, and a-melanocyte-stimulating hormone (aMSH) in bovine and rat intermediate pituitary has been studied.Rat anterior and intermediate pituitary rough endoplasmic reticulum/Golgi apparatus and cytosol fractions had similar amounts of acetyltransferase activity, but only the secretory granule fraction from the intermediate pituitary had what is believed to be the acetyltransferase activity specific for the formation of aMSH and a-Nacetyl-fi-endorphin in this tissue.Bovine intermediate pituitary secretory granules were used as the primary source of acetyltransferase activity.This activity, which used acetyl coenzyme A as the acetate donor, used ACTH-and P-endorphin-related peptides as the acetate acceptor.The bovine intermediate pituitary secretory granule-associated acetyltransferase activity was not membrane-bound and had a pH optimum of 7.0.ACTH(1-13)NHz, ACTH(1-39), aMSH, P-endorphin(1-31), and /3-endorphin(1-27) were among the effective peptide substrates.The products of the enzymatic reaction, using C3H]acetyl CoA as the acetate donor, were analyzed by chymotrypsin digestion followed by reverse-phase high pressure liquid chromatography; at least 75% of the radioactivity was associated with the NHz terminus of the peptide substrates.ACTH(18-39), lysine-rich histones, ACTH(l-Z)[Ser-Tyr], ACTH(l1-24), a-N,O-diacetyl-aMSH, and a-Nacetylated /3-endorphin-related molecules were not acetylated.Ser-Tyr, ACTH(l1-24), and a-N,O-diacetyl-aMSH inhibited the NH2-terminal acetylation of ACTH(1-13)NH2.Steady state kinetic experiments indicated that the K,,, value for acetyl CoA was 8 ~L M and the K,,, values for the peptide substrates ranged from 11 to 286 pM.The addition of a-N-acetyl-P-endorphin(1-27) inhibited the NHz-terminal acetylation of P-endorphin(1-27), ACTH(1-18), and ACTH(1-39); the inhibition of the acetylation of ACTH(1-18) by a-N-acetyl-/?endorphin(1-27) was competitive.These results s u g gest that the same enzyme NHz terminally acetylates both ACTH-and /I-endorphin-related peptides in the bovine intermediate pituitary.
Source: Journal of Biological Chemistry
Author: C C Glembotski
Institutions: University of Colorado Health, University of Colorado Denver
Language: English
Subfield: Cellular and Molecular Neuroscience
Field: Neuroscience
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,430
OpenAlex ID: W1929236293