The DegP protein, functioning as both chaperone and protease, plays a critical role in degrading and removing denatured or damaged proteins in the cellular envelope during heat shock and other stresses. So far, several proteins have been identified as its natural targets. A carboxyle-terminal peptide derived from the PapG pilus, one of the in vivo substrates for DegP, has been shown to activate the protease. Nevertheless, neither the details nor the physiological implications of such activation have been studied. The evidence that DegP undergoes conformational changes upon binding the peptide derived from C-terminal sequence of pilus subunit PapG has been presented. It demonstrated that upon binding this peptide, detectable changes can be observed for both secondary and tertiary structures of DegP, as examined by CD spectroscopy. Gel filtration and dynamic light scattering analysis also revealed that the size of DegP becomes smaller to a minor extent. Moreover, both the hydrophobic surfaces and catalytic sites of DegP were found to expose slightly in the presence of the peptide. Upon peptide binding, a less flexible and more rigid conformation of DegP was obtained as analyzed by fluorescence anisotropy. The physiological implications of these observations for DegP are discussed.