Purification of the Neisseria meningitidis transferrin binding protein-2 (TBP2) to homogeneity using column chromatography. 1993

L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, Spain.

A method for purifying TBP2 from N. meningitidis has been developed using affinity chromatography on Tf-agarose followed by ion exchange chromatography; the Tf-binding activity of fractions was evaluated by a dot-blot technique. This method allowed the purification of the TBP2 in milder conditions than those used previously and to a high degree of homogeneity as was demonstrated by Coomassie brilliant blue or Silver staining after SDS-PAGE electrophoresis. The SDS-PAGE profile of the material obtained in the Tf-agarose affinity chromatography step shows only two detectable proteins, identified as the TBP1 and the TBP2, with a small amount of contamination. Passage through a MonoQ HR anion exchange column, allowed the isolation of TBP2 in the absence of TBP1. Our results demonstrate the conservation of the antigenic reactivity of this protein, which produces monospecific serum with the antibodies elicited reacting exclusively with the TBP2 in whole outer membrane vesicles.

UI MeSH Term Description Entries
D007501 Iron A metallic element with atomic symbol Fe, atomic number 26, and atomic weight 55.85. It is an essential constituent of HEMOGLOBINS; CYTOCHROMES; and IRON-BINDING PROTEINS. It plays a role in cellular redox reactions and in the transport of OXYGEN. Iron-56,Iron 56
D009345 Neisseria meningitidis A species of gram-negative, aerobic BACTERIA. It is a commensal and pathogen only of humans, and can be carried asymptomatically in the NASOPHARYNX. When found in cerebrospinal fluid it is the causative agent of cerebrospinal meningitis (MENINGITIS, MENINGOCOCCAL). It is also found in venereal discharges and blood. There are at least 13 serogroups based on antigenic differences in the capsular polysaccharides; the ones causing most meningitis infections being A, B, C, Y, and W-135. Each serogroup can be further classified by serotype, serosubtype, and immunotype. Diplokokkus intracellularis meningitidis,Meningococcus,Micrococcus intracellularis,Micrococcus meningitidis,Micrococcus meningitidis cerebrospinalis,Neisseria weichselbaumii
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
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D002852 Chromatography, Ion Exchange Separation technique in which the stationary phase consists of ion exchange resins. The resins contain loosely held small ions that easily exchange places with other small ions of like charge present in solutions washed over the resins. Chromatography, Ion-Exchange,Ion-Exchange Chromatography,Chromatographies, Ion Exchange,Chromatographies, Ion-Exchange,Ion Exchange Chromatographies,Ion Exchange Chromatography,Ion-Exchange Chromatographies
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
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D014168 Transferrin An iron-binding beta1-globulin that is synthesized in the LIVER and secreted into the blood. It plays a central role in the transport of IRON throughout the circulation. A variety of transferrin isoforms exist in humans, including some that are considered markers for specific disease states. Siderophilin,Isotransferrin,Monoferric Transferrins,Serotransferrin,Transferrin B,Transferrin C,beta 2-Transferrin,beta-1 Metal-Binding Globulin,tau-Transferrin,Globulin, beta-1 Metal-Binding,Metal-Binding Globulin, beta-1,Transferrins, Monoferric,beta 1 Metal Binding Globulin,beta 2 Transferrin,tau Transferrin
D033862 Iron-Binding Proteins Proteins that specifically bind to IRON. Iron-Binding Protein,Iron Binding Protein,Iron Binding Proteins,Protein, Iron-Binding
D033863 Transferrin-Binding Proteins A class of carrier proteins that bind to TRANSFERRIN. Many strains of pathogenic bacteria utilize transferrin-binding proteins to acquire their supply of iron from serum. Transferrin Binding Proteins

Related Publications

L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
May 1995, Gene,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
August 1993, Gene,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
October 1994, Journal of bacteriology,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
January 2021, Turkish journal of chemistry,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
May 2005, Protein engineering, design & selection : PEDS,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
May 1990, FEMS microbiology letters,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
January 1996, Molecular microbiology,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
June 1992, Infection and immunity,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
September 2001, Infection and immunity,
L Ferrón, and C M Ferreirós, and M T Criado, and M P Andrade
March 2001, Infection and immunity,
Copied contents to your clipboard!