The electrophoretic mobility of gram-negative and gram-positive bacteria: an electrokinetic analysis.
The electrophoretic mobility (EPM) of a variety of Gram-negative and Gram-positive bacteria was measured with a Penkem S3000 analyser. Under standard growth conditions and neutral pH all cells displayed a negative EPM. The polysaccharide capsules of Escherichia coli strains K1, K5, K29 and K30 generated the highest EPM; to a lesser and varying degree O-antigens with charged groups and core lipopolysaccharides also contribute to the net EPM. Very little negative EPM was measured in suspension cultures of the gliding bacterium Cytophaga U67. No difference in the EPM was observed between rapidly growing and stationary-phase E. coli B. De-energization of the cell membranes by carbonyl cyanide m-chlorophenylhydrazone (CCCP) did not affect the EPM of wild-type and deep rough mutants of E. coli; and the EPM of Cytophaga U67 and Acholeplasma laidlawii remained unaltered by CCCP when measured in their respective growth media. Extrusion of filamentous bacteriophage f1 from cells of its host, E. coli A95, caused a shift to a higher negative EPM. We also measured a variety of Gram-positive strains, all of which displayed different EPMs. When membrane fractions of E. coli were adsorbed to latex spheres, characteristic differences between the EPM of beads coated with either inner or outer membrane were observed. The results suggest that the rapid EPM analysis is a useful tool to study the net electric charge of microorganisms and to examine changes of surface properties during interaction of cells with viruses, proteins (antibody) and charged antibiotics.
Bayer
M E
ME
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111.
Sloyer
J L
JL
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000942:Antigens, Bacterial; D002462:Cell Membrane / Q000032:analysis; D003090:Coliphages / Q000302:isolation & purification; D003470:Culture Media; D004586:Electrophoresis; D004926:Escherichia coli / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification; D006090:Gram-Negative Bacteria / Q000302:isolation & purification*; D006094:Gram-Positive Bacteria / Q000302:isolation & purification*; D019081:O Antigens
D000900:Anti-Bacterial Agents; D000942:Antigens, Bacterial; D003470:Culture Media; D019081:O Antigens