XENOPUS-LAEVIS SKIN ARG-XAA-VAL-ARG-GLY-ENDOPROTEASE - A HIGHLY SPECIFIC PROTEASE CLEAVING AFTER A SINGLE ARGININE OF A CONSENSUS SEQUENCE OF PEPTIDE-HORMONE PRECURSORS
- 5 September 1989
- journal article
- research article
- Vol. 264 (25) , 14609-14612
Abstract
Comparison of the precursor sequence for several peptide hormones of Xenopus laevis skin revealed a consensus sequence around a single arginine cleavage site which is 100% conserved on four residues Arg-Xaa-Val-Arg-Gly (RXVRG). A tetradecapeptide substrate (Asp-Val-Asp-Glu-Arg-Asp-Val-Arg-Gly-Phe-Ala-Ser-Phe-Leu-NH2) was used as a probe to purify and characterize the putative processing endoprotease. A hydrophobic enzyme was purified at least 9000-fold from Xenopus skin exudate by a four-step procedure. This highly specific activity cleaves the Arg-Gly bond and has no effect on the Arg-Xaa bond. It was strongly inhibited by divalent ion chelators, moderately by phenylmethylsulfonyl fluoride, aprotinin, and 1-tosylamide-2-phenylethyl chloromethyl ketone, but was insensitive to soybean trypsin inhibitor. Tetradecapeptide derivatives selectively modified on each of the amino acids of the consensus sequence demonstrated the relevance of this conserved pattern to endoprotease action. This enzyme, which we refer to as RXVRG-endoprotease, is proposed to be involved in the post-translational processing of pro-caerulein, promagainin, pro-xenopsin, pro-glycyl-leucine amide, and pro-levitide of X. laevis skin secretory granules.This publication has 10 references indexed in Scilit:
- Proocytocin neurophysin convertase from bovine neurohypophysis and corpus luteum secretory granules: complete purification, structure-function relationships, and competitive inhibitorBiochemistry, 1989
- Yeast KEX2 gene encodes an endopeptidase homologous to subtilisin-like serine proteasesBiochemical and Biophysical Research Communications, 1988
- Synthetic peptide substrates as models to study a pro-ocytocin/neurophysin converting enzymeJournal of Chromatography A, 1988
- Yeast KEX1 gene encodes a putative protease with a carboxypeptidase B-like function involved in killer toxin and α-factor precursor processingCell, 1987
- Expression of porcine pro-opiomelanocortin cDNA in heterologous monkey kidney cells. Biosynthesis and secretion of the prohormone without processing.Journal of Biological Chemistry, 1987
- Skin peptides in Xenopus laevis: morphological requirements for precursor processing in developing and regenerating granular skin glands.The Journal of cell biology, 1986
- Solid phase synthesis of somatostatin-28 II. A new biologically active octacosapeptide from anglerfish pancreatic isletsBiochemical and Biophysical Research Communications, 1986
- Proteolytic conversion of [Met]enkephalin-Arg6-Gly7-Leu8 by brain synaptic membranes. Characterization of formed peptides and mechanism of proteolysis.Journal of Biological Chemistry, 1985
- Biosynthesis of thyrotropin releasing hormone in the skin of Xenopus laevis: partial sequence of the precursor deduced from cloned cDNA.The EMBO Journal, 1984
- Mechanism of action of thrombin on fibrinogen. Kinetic evidence for involvement of aspartic acid at position P10Biochemistry, 1983