Gradient, polyacrylamide gel electrophoresis of proteins from cytotoxic mycoplasma membranes

Membranes of Mycoplasma pneumoniae were prepared following several different cell lysis procedures (freeze-thaw, french press, digitonin, sonication, osmotic shock, pH shock). All possessed some degree of toxicity for ciliated epithelial cells, but the french press and freeze-thaw processes were optimal in terms of lysis efficiency, membrane yield, and cytotoxicity. Membrane proteins from virulent and attenuated mycoplasmas were electrophoresed with a new, high resolution electrophoresis technique (gradient polyacrylamide gels), and strain differences were detectable among the 50+ discernible bands.