Biomaterial Database

Protein modification of glutamine transporters in SV40-transformed hepatocytes and immunodetection of proteins associated with hepatic system N transport activity. 1994

B K Tamarappoo, and H P Singh, and M S Kilberg

Department of Medicine, University of Florida College of Medicine, Gainesville 32610.

The secondary active transport of glutamine into hepatocytes is mediated by the sodium-dependent System N amino acid transporter. In SV40-transformed fetal or adult hepatocytes, System N activity was increased by culturing the cells at the growth-permissive temperature of 33 degrees C compared with 40 degrees C. The sulfhydryl specific protein modification reagent p-chloromercuribenzene sulfonate (PCMBS) inactivated all of System N activity in fetal cells, whereas in the adult cells PCMBS inactivated only the elevated System N activity expressed at 33 degrees C; basal System N activity remained unaffected. Also, the inactivation of transport in the adult cells could be blocked by the presence of the substrate glutamine, whereas that in the fetal cells was completely resistant to protection. To characterize the System N transport, murine monoclonal antibodies System N transporter, murine monoclonal antibodies were generated. Two hybridomas (3E1-2 and 1E7-3) produced antibodies that inhibit System N activity in hepatocytes and can immunoprecipitate the activity from a solubilized protein fraction. Based on two-dimensional PAGE, the molecular size of the carrier protein has been shown to be approximately 100 kDa.

UI	MeSH Term	Description	Entries
D008099	Liver	A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.	Livers
D011233	Precipitin Tests	Serologic tests in which a positive reaction manifested by visible CHEMICAL PRECIPITATION occurs when a soluble ANTIGEN reacts with its precipitins, i.e., ANTIBODIES that can form a precipitate.	Precipitin Test,Test, Precipitin,Tests, Precipitin
D002352	Carrier Proteins	Proteins that bind or transport specific substances in the blood, within the cell, or across cell membranes.	Binding Proteins,Carrier Protein,Transport Protein,Transport Proteins,Binding Protein,Protein, Carrier,Proteins, Carrier
D002461	Cell Line, Transformed	Eukaryotic cell line obtained in a quiescent or stationary phase which undergoes conversion to a state of unregulated growth in culture, resembling an in vitro tumor. It occurs spontaneously or through interaction with viruses, oncogenes, radiation, or drugs/chemicals.	Transformed Cell Line,Cell Lines, Transformed,Transformed Cell Lines
D004591	Electrophoresis, Polyacrylamide Gel	Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.	Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D005333	Fetus	The unborn young of a viviparous mammal, in the postembryonic period, after the major structures have been outlined. In humans, the unborn young from the end of the eighth week after CONCEPTION until BIRTH, as distinguished from the earlier EMBRYO, MAMMALIAN.	Fetal Structures,Fetal Tissue,Fetuses,Mummified Fetus,Retained Fetus,Fetal Structure,Fetal Tissues,Fetus, Mummified,Fetus, Retained,Structure, Fetal,Structures, Fetal,Tissue, Fetal,Tissues, Fetal
D005973	Glutamine	A non-essential amino acid present abundantly throughout the body and is involved in many metabolic processes. It is synthesized from GLUTAMIC ACID and AMMONIA. It is the principal carrier of NITROGEN in the body and is an important energy source for many cells.	D-Glutamine,L-Glutamine,D Glutamine,L Glutamine
D006825	Hybridomas	Cells artificially created by fusion of activated lymphocytes with neoplastic cells. The resulting hybrid cells are cloned and produce pure MONOCLONAL ANTIBODIES or T-cell products, identical to those produced by the immunologically competent parent cell.	Hybridoma
D000375	Aging	The gradual irreversible changes in structure and function of an organism that occur as a result of the passage of time.	Senescence,Aging, Biological,Biological Aging
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