Striated fibers of the rho form of Mycoplasma: in vitro reassembly, composition, and structure.
The rho-form of Mycoplasma contains a striated, axial fiber and associated terminal structure. The presence of this organelle was correlated with the synthesis of two proteins, A and B, of molecular weights of approximately 85,000 and 26,000, respectively, each accounting for about 10% of the total cell protein. Their amino acid compositions showed them to have distinct polypeptide chains. After osmotic lysis of rho-form cells the organelles disappeared; protein A accompanied the membrane fraction, whereas protein B was partly released in soluble form. After lysis by Nonidet P-40 in a medium composed of 4 M glycerol, 50 mM phosphate, and 10 mM MgSO4 at pH 6 (GPM-6), the organelles were preserved and released with ultrastructure unchanged. Protein A was recovered in the soluble fraction and protein B in the particulate (crude fiber) fraction. Treatment of the crude fiber fraction with 0.5 M NaCl in GPM-6 or with a solution containing 4 M glycerol, 10 mM morpholinoethanesulfonate, and 1 mM ethylenediaminetetraacetate at pH 7.0 caused the fibers to disassemble into subunits. By subsequent changes in the ionic conditions and temperature it was possible to cause the subunits to reassemble into ordered aggregates having the same ultrastructure as the native rho-fibers. The optimum temperature for reassembly in the presence of 4 M glycerol was 37 C, the optimum pH was 6.5 to 7.0, and the presence of Mg-2+, replaceable by Ca-2+, SR-2+, or Ba-2+, was essential. Protein B was the only protein detected in the purified, reconsituted fibers.
D000255:Adenosine Triphosphate; D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism; D001433:Bacteriolysis; D002458:Cell Fractionation; D002850:Chromatography, Gel; D003470:Culture Media; D004072:Digitonin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005990:Glycerol; D006160:Guanosine Triphosphate; D006863:Hydrogen-Ion Concentration; D008274:Magnesium; D008970:Molecular Weight; D009024:Morphogenesis; D009174:Mycoplasma / Q000032:analysis / Q000648:ultrastructure*; D009940:Organoids / Q000032:analysis / Q000648:ultrastructure*; D009995:Osmosis; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature
D000596:Amino Acids; D001426:Bacterial Proteins; D003470:Culture Media; D013501:Surface-Active Agents; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D008274:Magnesium; D004072:Digitonin; D005990:Glycerol