Identification and characterization of an insect homologue of the vertebrate Golgi apparatus protein 1 (MG-160/cysteine-rich fibroblast growth factor receptor/E-selectin ligand-1/latent transforming growth factor-beta complex protein-1) with a Golgi-specific monoclonal antibody.
A monoclonal antibody 14F10 was raised against Golgi fractions from Sf21 cells and selected as Golgi specific. Immunohistochemical stainings with the antibody localized the antigen in Golgi cisterns of the cells. The antigen was purified and shown to be a 130-K membrane protein with N-glycans and intrachain disulfide bonds. Amino acid sequencing of its peptide fragments revealed that the antigen contained homologous sequences to those encoded by CG7190 and CG7193 Drosophila melanogaster genes. No possible transmembrane domain existed in these deduced amino acid sequences, while one did in that encoded by CG7195, an adjacent gene to CG7193. Furthermore, 5' and 3' expression sequence tags of LD19434 had been mapped to CG7190 and a downstream region of CG7195, respectively. These findings supported that all of these genes actually composed a single gene, which encoded an orthologous protein to a vertebrate Golgi-resident protein, Golgi apparatus protein 1, also called cysteine-rich FGF receptor, E-selectin ligand-1, or latent TGF-beta complex protein-1. Our results suggested that the Golgi apparatus protein 1 played a critical role in the Golgi cisterns through the animal kingdom.
Kawano
Jun-ichi
J
Department of Anatomy, Miyazaki Medical College, Kiyotake, Miyazaki 889-1692, Japan. jkawano@post.miyazaki-med.ac.jp
Nakayama
Tatsuo
T
Kotani
Tomio
T
Matsubayashi
Hiroshi
H
Yamamoto
Masa-Toshi
MT
Suganuma
Tatsuo
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology; D002460:Cell Line; D029721:Drosophila Proteins / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000737:chemistry* / Q000276:immunology / Q000648:ultrastructure; D007150:Immunohistochemistry; D053779:Latent TGF-beta Binding Proteins; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011956:Receptors, Cell Surface / Q000235:genetics; D017468:Receptors, Fibroblast Growth Factor / Q000032:analysis* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012795:Sialoglycoproteins / Q000235:genetics; D018411:Spodoptera / Q000737:chemistry / Q000235:genetics; D014714:Vertebrates / Q000378:metabolism
D000911:Antibodies, Monoclonal; D029721:Drosophila Proteins; D053779:Latent TGF-beta Binding Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D017468:Receptors, Fibroblast Growth Factor; D012795:Sialoglycoproteins; C086711:cysteine-rich fibroblast growth factor receptor