Identification and characterization of a novel Golgi protein, golgin-67.
In the course of screening a lambdagt11 human leukemic T-cell cDNA expression library with an antibody specific to the mitotic target of Src, Sam68, we identified and cloned a cDNA encoding a novel protein with a predicted molecular mass of 51.4 kDa. Polyclonal antibodies raised to a His(6)-tagged construct of this protein, detected a approximately 67-kDa protein in immunoprecipitation experiments, and cytological studies showed that this protein localized to the Golgi complex, through colocalization experiments with specific Golgi markers. Therefore, we designated this protein golgin-67. Sequence analysis revealed that golgin-67 is a highly coiled-coil protein, with potential Cdc2 and Src kinase phosphorylation motifs. It has sequence homologies to other Golgi proteins, including the coatamer complex I vesicle docking protein, GM130. Structurally, golgin-67 resembles, golgin-84, an integral membrane Golgi protein with an N-terminal coiled-coil domain and a single C-terminal transmembrane domain. The C-terminal region of golgin-67, which contains a predicted transmembrane domain, was demonstrated to be essential for its Golgi localization.
Jakymiw
A
A
Cancer Biology Research Group, Department of Biochemistry and Molecular Biology, University of Calgary Medical Center, Calgary, Alberta T2N 4N1, Canada.
Raharjo
E
E
Rattner
J B
JB
Eystathioy
T
T
Chan
E K
EK
Fujita
D J
DJ
D000595:Amino Acid Sequence; D015415:Biomarkers; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000737:chemistry*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011487:Protein Conformation; D012333:RNA, Messenger / Q000378:metabolism; D013601:T-Lymphocytes / Q000737:chemistry; D013737:Testis / Q000737:chemistry
D015415:Biomarkers; D008565:Membrane Proteins; D012333:RNA, Messenger; C404952:golgin-67