title: A peptidyl-prolyl cis/trans-isomerase (cyclophilin G) in regulated secretory granules.
abstract: A 27-kDa protein (p27) in horseshoe crab hemocyte that cross-reacts with antiserum against a beta-glucan-sensitive protease zymogen was purified to homogeneity, and its cDNA was cloned. The 1.7-kilobase pair cDNA contains an open reading frame of 660 base pairs, encoding a 23-amino acid signal sequence followed by a mature protein of 197 residues. The sequence of p27 exhibits strong similarity to that of cyclophilin B, a peptidyl-prolyl cis/trans-isomerase. p27 exhibits isomerase activity with a kcat/Km of 0.18 microM-1 s-1 for a peptide substrate; this activity is inhibited by cyclosporin A but is not affected by FK506. Although the p27 precursor possesses an amino-terminal secretory hydrophobic signal sequence, unlike other cyclophilin B molecules, it lacks a conserved carboxyl-terminal endoplasmic reticulum retention signal and it contains a central 8-amino acid insertion. Although p27 is secreted into the culture media of transiently expressed COS cells, it is not detected in horseshoe crab hemolymph plasma but rather is localized to the hemocyte large granules, the regulated secretory granules that are exocytosed upon stimulation. These results indicate that p27 is a new peptidyl-prolyl cis/trans-isomerase in the regulated secretory granules, and is thus designated cyclophilin G. This first report of a cyclophilin homologue in the secretory granule of the horseshoe crab hemocyte suggests that such chaperon-like proteins may constitute a key quality control system for stored proteins in exocytotic granules.
authors.affiliation: Department of Molecular Biology, Graduate School of Medical Science, Kyushu University 33, Fukuoka 812-81, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Takaki
authors.forename: T
authors.initials: T
authors.lastname: Muta
authors.forename: S
authors.initials: S
authors.lastname: Iwanaga
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003594:Cytoplasmic Granules / Q000201:enzymology*; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D006434:Hemocytes / Q000201:enzymology; D006737:Horseshoe Crabs; D008969:Molecular Sequence Data; D019696:Peptidylprolyl Isomerase / Q000737:chemistry / Q000302:isolation & purification*; D016415:Sequence Alignment
chemical_list: D018076:DNA, Complementary; D019696:Peptidylprolyl Isomerase