Overproduction, purification and characterization of the HPB12-L24 ribosomal protein of Bacillus subtilis

• Published in: FEMS microbiology letters 1996-11, Vol.145 (1), p.41-48
• Main Authors: Zouine, Mohamed, Beloin, Christophe, Deneubourg, Anne-Marie, Hirschbein, Luisa, Le Hegarat, Françoise
• Format: Article
• Language: English
• Subjects: Ammonium; Ammonium sulfate; Antibody Specificity; Bacillus subtilis; Bacillus subtilis - genetics; Bacillus subtilis - metabolism; Bacterial Proteins; Bacteriological Techniques; Cation exchanging; Cation-exchange chromatography; Cell morphology; Cytology; Deoxyribonucleic acid; DNA; DNA, Bacterial; DNA, Bacterial - metabolism; DNA-Binding Proteins; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - isolation & purification; E coli; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Escherichia coli - growth & development; Escherichia coli - immunology; Gel retardation; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Bacterial - physiology; Heat exchange; Histone‐like protein; L24 ribosomal protein; Life Sciences; Microbiology; Morphology; Nucleoid; Nucleoids; Overproduction; Protein purification; Proteins; Purification; Recombinant Proteins; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Ribosomal Proteins; Ribosomal Proteins - chemistry; Ribosomal Proteins - genetics; Ribosomal Proteins - isolation & purification; Thermal stability
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.1996.tb08554.x

Abstract HPB12-L24 was previously described as a bifunctional histone-like and ribosomal protein in Bacillus subtilis. In order to confirm the identity of HPB12 and L24, and to study the properties of this protein, the rplX gene of B. subtilis encoding L24 has been overexpressed in Escherichia coli by an efficient protein overproduction system. A simple and rapid purification scheme using ammonium sulfate precipitation and cation-exchange chromatography is presented. 10 mg of pure L24 per g of Escherichia coli cells were obtained. The purified recombinant protein L24 is heat-stable, acid-soluble and binds preferentially supercoiled DNA like protein HPB12. These results confirm the identity of HPB12 and L24. Overexpression of rplX led to gross alterations of cell morphology and to an abnormal shape of nucleoids.