Molecular cloning, characterization, and dynamics of rat formiminotransferase cyclodeaminase, a Golgi-associated 58-kDa protein. 1998

Y S Gao, and C Alvarez, and D S Nelson, and E Sztul
Department of Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.

A peripherally associated 58-kDa Golgi protein (58K) of unknown function has been previously described (Bloom, G. S., and Brashear, T. A. (1989) J. Biol. Chem. 264, 16083-16092). To molecularly characterize 58K, we used a monoclonal anti-58K antibody (monoclonal antibody 58K-9) to screen a rat liver cDNA expression library. Positive clones were isolated, characterized, and partially sequenced. The obtained sequences show a high level of identity with sequences of porcine formiminotransferase cyclodeaminase (FTCD), suggesting that 58K is rat FTCD. Rat FTCD is structurally similar to porcine FTCD, a metabolic enzyme involved in conversion of histidine to glutamic acid, and exists in dimeric, tetrameric, and octameric complexes resistant to proteolysis. To define parameters of FTCD association with the Golgi, comparison of its behavior with various Golgi and ER-to-Golgi intermediate compartment marker proteins was examined under specific conditions. The results show that extraction parameters of FTCD are similar to those of GM130, a tightly associated Golgi matrix protein. FTCD appears to be a dynamic component of the Golgi, and a proportion of FTCD molecules cycle between the Golgi and earlier compartments of the secretory pathway. FTCD remains associated with Golgi fragments during microtubule disruption and is not released into cytosol during brefeldin A treatment. Instead, FTCD relocates from the Golgi, but the time course of its redistribution is distinct from that of mannosidase II relocation. FTCD is already dispersed into small punctate structures at a time when mannosidase II is still largely localized to Golgi structures. FTCD is not observed in tubules originating from the Golgi and containing mannosidase II. Instead, it appears to redistribute in small vesicles arranged in a linear "pearls on a string" pattern. These results suggest that FTCD relocation is temporally and spatially distinct from mannosidase II relocation and that FTCD provides a novel marker to study Golgi dynamics.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D002451 Cell Compartmentation A partitioning within cells due to the selectively permeable membranes which enclose each of the separate parts, e.g., mitochondria, lysosomes, etc. Cell Compartmentations,Compartmentation, Cell,Compartmentations, Cell
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D004721 Endoplasmic Reticulum A system of cisternae in the CYTOPLASM of many cells. In places the endoplasmic reticulum is continuous with the plasma membrane (CELL MEMBRANE) or outer membrane of the nuclear envelope. If the outer surfaces of the endoplasmic reticulum membranes are coated with ribosomes, the endoplasmic reticulum is said to be rough-surfaced (ENDOPLASMIC RETICULUM, ROUGH); otherwise it is said to be smooth-surfaced (ENDOPLASMIC RETICULUM, SMOOTH). (King & Stansfield, A Dictionary of Genetics, 4th ed) Ergastoplasm,Reticulum, Endoplasmic
D006056 Golgi Apparatus A stack of flattened vesicles that functions in posttranslational processing and sorting of proteins, receiving them from the rough ENDOPLASMIC RETICULUM and directing them to secretory vesicles, LYSOSOMES, or the CELL MEMBRANE. The movement of proteins takes place by transfer vesicles that bud off from the rough endoplasmic reticulum or Golgi apparatus and fuse with the Golgi, lysosomes or cell membrane. (From Glick, Glossary of Biochemistry and Molecular Biology, 1990) Golgi Complex,Apparatus, Golgi,Complex, Golgi
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000642 Ammonia-Lyases Enzymes that catalyze the formation of a carbon-carbon double bond by the elimination of AMMONIA. EC 4.3.1. Ammonia Lyase,Ammonia-Lyase,Ammonia Lyases,Lyase, Ammonia
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013552 Swine Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA). Phacochoerus,Pigs,Suidae,Warthogs,Wart Hogs,Hog, Wart,Hogs, Wart,Wart Hog

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