title: Purification, characterization, and molecular cloning of an outer layer protein of carp fertilization envelope.
abstract: An outer layer protein of carp fertilization envelope (FE), FEO-1, was purified from carp oocytes. The cDNAs encoding FEO-1 were cloned. The mature protein of FEO-1 is 21 kDa in molecular weight and contains 177 amino acid residues whose sequence has 58% identity to the outer layer protein of chick vitelline membrane. In situ hybridization and immunocytochemistry show that FEO-1 is expressed in oocytes and liver. In oocytes, FEO-1 is stored in the cortical granules. During cortical reaction, it is exocytosed to the perivitelline space and then gradually added to the outer layer of FE (FE(o)). FEO-1 first appears as discrete deposits along FE(o), then merges to form a continuous layer. The thickness of FE(o) increases as cortical reaction proceeds. In addition to FEO-1, FE(o) contains cystatin, fibroin-like substance (FLS), and cathepsin-like substance (CLS) as well. They are stored in the cortical granules and are exocytosed to FE(o) simultaneously with FEO-1 during cortical reaction. In FE(o), FEO-1 is present in monomer form and can be completely extracted by sodium dodecyl sulfate (SDS)-mercaptoethanol (MSH). On the other hand, the cystatin, FLS, and CLS present in FE(o)are cross-linked together. They are partially extracted by SDS-MSH but can be completely extracted by guanidium thiocyanate-lauroylsarcosine.
authors.affiliation: Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
authors.forename: Y S
authors.initials: YS
authors.lastname: Chang
authors.forename: L F
authors.initials: LF
authors.lastname: Lu
authors.forename: C Y
authors.initials: CY
authors.lastname: Lai
authors.forename: Y H
authors.initials: YH
authors.lastname: Kou
authors.forename: F L
authors.initials: FL
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002347:Carps / Q000235:genetics* / Q000378:metabolism*; D002403:Cathepsins / Q000032:analysis; D002642:Chick Embryo; D002823:Chorion / Q000378:metabolism*; D003001:Cloning, Molecular; D015891:Cystatins / Q000032:analysis; D018076:DNA, Complementary / Q000032:analysis; D004527:Egg Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005089:Exocytosis; D005349:Fibroins / Q000032:analysis; D029941:Fish Proteins; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009865:Oocytes / Q000737:chemistry*; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D015891:Cystatins; D018076:DNA, Complementary; D004527:Egg Proteins; C122031:FEO-1 protein, Cyprinus carpio; D029941:Fish Proteins; D011994:Recombinant Proteins; D005349:Fibroins; D002403:Cathepsins