Mycoplasma pneumoniae protein P200 was localized to the terminal organelle, which functions in cytadherence and gliding motility. The loss of P200 had no impact on binding to erythrocytes and A549 cells but resulted in impaired gliding motility and colonization of differentiated bronchial epithelium. Thus, gliding may be necessary to overcome mucociliary clearance.Mycoplasma pneumoniae causes tracheobronchitis, bronchopneumonia, and a variety of extrapulmonary manifestations in humans (35). A distinct polar structure (5, 18) mediates host cell attachment (cytadherence) by this novel cell wall-less prokaryote. Electron micrographs of experimentally infected human and animal cells, as well as cells collected from natural M. pneumoniae infections, reveal an intimate relationship between this terminal organelle and the epithelial cell surface (7). The terminal organelle also mediates gliding motility (16,29), which may facilitate mycoplasma traversal of the airway surface liquid to allow access to host receptors on the apical surface of respiratory epithelium and refuge from the turbulent mucociliary escalator.