Processing enzyme specificity is a consequence of pro-hormone precursor protein conformation.
Peptide-hormones are synthesized as higher molecular weight, precursor proteins which must initially undergo limited endoproteolysis to yield the bioactive peptide(s). The ability of two different endoproteinases, gonadotropin-associated peptide (GAP)-releasing enzyme and atrial granule serine proteinase (which are likely to be the physiologically relevant processing enzymes of bovine hypothalamic pro-gonadotropin-releasing hormone/gonadotropin-associated peptide and bovine pro-atrial natriuretic factor precursor proteins, respectively), to act at their own recognition sequences within their relevant pro-hormone proteins has now been contrasted with their ability to act at the recognition sequence for the alternate enzyme or to act at their own recognition sequence when it is placed within the protein framework of the alternate precursor protein. The results show that each enzyme acts with specificity at its own recognition sequence even when it is placed within the framework of the alternate pro-hormone. However, the enzymes fail to act (or act in a non-specific manner) at the alternate recognition sequence even if it is placed within the peptide framework of its own pro-hormone protein. Thus, despite the fact that both recognition sequences are similar in sequence and residue composition and that both contain a doublet of basic amino acids, it appears that sequence and the local conformation assumed by the processing site within the pro-hormone protein are essential for each endoproteinase to act with fidelity. As part of our continuing work, we now also report several newly determined physicochemical properties of hypothalamic GAP-releasing enzyme, the processing enzyme of pro-gonadotropin-releasing hormone/GAP protein.
Rangaraju
N S
NS
Department of Biochemistry, Virginia Commonwealth University, Richmond 23298-0614.
Harris
R B
RB
D000595:Amino Acid Sequence; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000737:chemistry; D002417:Cattle; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D006062:Gonadotropins / Q000737:chemistry; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity
D006062:Gonadotropins; D010455:Peptides; D011498:Protein Precursors; D009320:Atrial Natriuretic Factor; D010450:Endopeptidases; C052441:gonadotropin-associated-peptide releasing enzyme