title: Overproduction, purification and characterization of the HPB12-L24 ribosomal protein of Bacillus subtilis.
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.
authors.affiliation: Institut de Génétique et Microbiologie, Université Paris-Sud, Orsay, France.
authors.forename: M
authors.initials: M
authors.lastname: Zouine
authors.forename: C
authors.initials: C
authors.lastname: Beloin
authors.forename: A M
authors.initials: AM
authors.lastname: Deneubourg
authors.forename: L
authors.initials: L
authors.lastname: Hirschbein
authors.forename: F
authors.initials: F
authors.lastname: Le Hegarat
mesh_terms: D000918:Antibody Specificity; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins; D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000254:growth & development / Q000276:immunology; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C085226:HPB12 protein, Bacillus subtilis; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C036298:ribosomal protein L24