Highly selective extraction of spiralin from the Spiroplasma citri cell membrane with alkyl-N-sulfobetaines.
The extraction of proteins from the membrane of the mollicute (mycoplasma) Spiroplasma citri by sodium N-dodecyl-N,N-dimethyl-3-amino-1-propane sulfonate (SB12) and sodium N-tetradecyl-N,N-dimethyl-3-amino-1-propane sulfonate (SB14) was studied with electrophoretic methods. The membranes were prepared by osmotic lysis of the cells and depleted of the bulk of extrinsic proteins. It was possible to extract up to 35 and 45% of membrane proteins with SB12 and SB14, respectively. Maximal yield was obtained in both cases with detergent concentrations greater than or equal to 5 mumoles/mg of membrane protein. Spiralin, the major protein in the S. citri membrane, was highly selectively solubilized without the loss of antigenicity, with a yield of about 90% with SB12 and close to 100% with SB14, for a detergent concentration greater than or equal to 0.2 M. The degree of selectivity in favour of spiralin was higher with SB12 (purity approximately equal to 70%) than with SB14 (purity approximately equal to 50%). Treatment of the S. citri membrane with high concentrations of SB12 is a simple and fast procedure for partial purification of spiralin. This example shows that, in some cases, it should be possible to modulate the selectivity of the extraction of membrane proteins simply by varying the relative concentration of detergent.
D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification*; D002462:Cell Membrane / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000302:isolation & purification; D008722:Methods; D010455:Peptides / Q000302:isolation & purification; D000644:Quaternary Ammonium Compounds; D013149:Spiroplasma / Q000032:analysis*
D001425:Bacterial Outer Membrane Proteins; D008565:Membrane Proteins; D010455:Peptides; D000644:Quaternary Ammonium Compounds; C028588:spiralin; C034730:zwittergent 3-12; C048256:zwittergent 3-14