Presence of a particulate thyrotropin-releasing hormone-degrading pyroglutamate aminopeptidase activity in rat liver.
The data of this work describe the presence in the rat liver of a thyrotropin-releasing hormone (TRH)-degrading particulate metal-containing pyroglutamate aminopeptidase of high molecular weight. Following the fractionation of liver homogenate in 0.25 M sucrose, solubilization of the particulate fraction with papain and gel filtration on ACA34, an enzyme activity was detected which converts TRH into pyroglutamic acid and histidyl-proline diketopiperazine (cyclo[His-Pro]; cHP). [L-Histidine-2,5-3H]TRH and [L-proline-2,3-3H]TRH were used as a tracer. Products formed were separated by thin-layer chromatography, localized and quantified by scanning for radioactivity. Enzyme activities were tested using preferential site-directed inhibitors. Particulate pyroglutamate aminopeptidase activity was found to be sensitive to chelating agents. The physicochemical properties of this particulate aminopeptidase were distinct from the soluble pyroglutamate aminopeptidase from the same source. The particulate enzyme shared several similarities with particulate pyroglutamate aminopeptidase from adenohypophysis, brain and with serum enzyme, reported to have narrower specificity for TRH compared to the soluble pyroglutamate aminopeptidase from several tissues. The Km of the gel-filtrated enzyme is 27 microM and the specific activity 306 pmol.min-1.mg protein-1. Although no definite role has been established for this enzyme, it might be a potential determinant of cHP concentrations in liver. Furthermore, cHP is known to possess biological activities and specific binding sites in liver membranes. One of the major sites for TRH breakdown in vivo, the liver, probably represents a target tissue for TRH, especially for pancreatic TRH, and the particulate enzyme involved in the conversion of TRH into cHP may assume a specific function.
Scharfmann
R
R
INSERM U.30, Hôpital des Enfants Malades, Paris, France.
Morgat
J L
JL
Aratan-Spire
S
S
D000626:Aminopeptidases / Q000378:metabolism*; D000818:Animals; D008099:Liver / Q000201:enzymology* / Q000378:metabolism*; D008297:Male; D010456:Peptides, Cyclic / Q000096:biosynthesis; D010879:Piperazines / Q000096:biosynthesis; D011751:Pyroglutamyl-Peptidase I / Q000378:metabolism*; D011761:Pyrrolidonecarboxylic Acid / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism; D013973:Thyrotropin-Releasing Hormone / Q000378:metabolism*
D010456:Peptides, Cyclic; D010879:Piperazines; D013973:Thyrotropin-Releasing Hormone; D000626:Aminopeptidases; D011751:Pyroglutamyl-Peptidase I; C015924:histidyl-proline diketopiperazine; D011761:Pyrrolidonecarboxylic Acid