Haemophilus influenzae penicillin-binding proteins 1a and 3 possess distinct and opposite temperature-modulated penicillin-binding activities.

Upon investigation of penicillin-binding proteins (PBPs) in Haemophilus influenzae strains, five H. influenzae and seven other Haemophilus strains were tested for whole-cell penicillin binding at either 37 or 42 degrees C. Binding of [35S]penicillin G to H. influenzae PBPs 3a and 3b was drastically reduced at 42 degrees C, while PBP 1a showed a temperature-modulated increase in penicillin binding. Further investigation revealed that growth at 42 degrees C causes altered electrophoretic mobility of PBP 3a on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and that cell labeling performed at 42 degrees C showed the differential penicillin binding to target proteins. All Haemophilus spp. tested showed a similar temperature modulation of penicillin binding. Growth measurement and cell viability studies performed at 42 degrees C permitted correlation of PBP 3 temperature sensitivity to H. influenzae resistance to moxalactam at 42 degrees C, and the probable correlation of PBP 1a increased penicillin binding to the more rapid antibacterial activity of penicillin G against H. influenzae at 42 degrees C. Microscopic examination of Haemophilus cells grown at 42 degrees C revealed filamentous cell formation, supporting a role of PBP 3 in the septation process. Results of this study demonstrate that wild-type H. influenzae strains (and possibly all other Haemophilus spp.) possess PBPs 1a and 3, which have distinct and opposite temperature-modulated penicillin-binding activities.