Hydrodynamic properties and immunological identification of the sodium- and chloride-coupled glycine transporter. 1993

B López-Corcuera, and R Alcántara, and J Vázquez, and C Aragón
Departamento de Biología Molecular, Facultad de Ciencias, Universidad Autónoma, Madrid, Spain.

We have recently reported the purification of the native sodium- and chloride-coupled glycine transporter from pig brain stem (López-Corcuera, B. Vázquez, J., and Aragón, C. (1991) J. Biol. Chem. 266, 24809-24814). This preparation is essentially homogeneous and contains a unique 100-kDa polypeptide based on electrophoretic migration under denaturing conditions. In this paper we report the hydrodynamic characterization of the native transporter, solubilized in two different detergents, as well as the immunological identification of the protein. On the basis of results obtained from size-exclusion chromatography, we calculated the Stokes radii of transporter 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS) and transporter-cholate complexes to be 5.5 and 6.0 nm, respectively. In addition, from H2O/D2O sucrose density gradient sedimentation analysis, we calculated the molecular weight of protein-detergent complexes to be 115,000 and 160,000 in CHAPS and cholate, respectively, and the molecular weight of the protein moiety as 86,000. Finally, polyclonal antibodies raised against the 100-kDa polypeptide were found to immunoprecipitate specifically glycine transport activity. Taken together, the results reported herein corroborate the identity of the 100-kDa band as the sodium- and chloride-coupled glycine transporter and also suggest that in its native state the transporter is a monomeric protein.

UI MeSH Term Description Entries
D007106 Immune Sera Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen. Antisera,Immune Serums,Sera, Immune,Serums, Immune
D007163 Immunosorbent Techniques Techniques for removal by adsorption and subsequent elution of a specific antibody or antigen using an immunosorbent containing the homologous antigen or antibody. Immunoadsorbent Techniques,Immunoadsorbent Technics,Immunosorbent Technics,Immunoadsorbent Technic,Immunoadsorbent Technique,Immunosorbent Technic,Immunosorbent Technique,Technic, Immunoadsorbent,Technic, Immunosorbent,Technics, Immunoadsorbent,Technics, Immunosorbent,Technique, Immunoadsorbent,Technique, Immunosorbent,Techniques, Immunoadsorbent,Techniques, Immunosorbent
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D001933 Brain Stem The part of the brain that connects the CEREBRAL HEMISPHERES with the SPINAL CORD. It consists of the MESENCEPHALON; PONS; and MEDULLA OBLONGATA. Brainstem,Truncus Cerebri,Brain Stems,Brainstems,Cerebri, Truncus,Cerebrus, Truncus,Truncus Cerebrus
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
D002499 Centrifugation, Density Gradient Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Centrifugations, Density Gradient,Density Gradient Centrifugation,Density Gradient Centrifugations,Gradient Centrifugation, Density,Gradient Centrifugations, Density
D002627 Chemistry, Physical The study of CHEMICAL PHENOMENA and processes in terms of the underlying PHYSICAL PHENOMENA and processes. Physical Chemistry,Chemistries, Physical,Physical Chemistries
D002712 Chlorides Inorganic compounds derived from hydrochloric acid that contain the Cl- ion. Chloride,Chloride Ion Level,Ion Level, Chloride,Level, Chloride Ion
D002793 Cholic Acids The 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholanic acid family of bile acids in man, usually conjugated with glycine or taurine. They act as detergents to solubilize fats for intestinal absorption, are reabsorbed by the small intestine, and are used as cholagogues and choleretics. Cholalic Acids,Acids, Cholalic,Acids, Cholic
D002850 Chromatography, Gel Chromatography on non-ionic gels without regard to the mechanism of solute discrimination. Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography

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