Energetic rationale for an unexpected and abrupt reversal of guanidinium chloride-induced unfolding of peptide deformylase.
Peptide deformylase (PDF) catalyzes the removal of formyl group from the N-terminal methionine residues of nascent proteins in prokaryotes, and this enzyme is a high priority target for antibiotic design. In pursuit of delineating the structural-functional features of Escherichia coli PDF (EcPDF), we investigated the mechanistic pathway for the guanidinium chloride (GdmCl)-induced unfolding of the enzyme by monitoring the secondary structural changes via CD spectroscopy. The experimental data revealed that EcPDF is a highly stable enzyme, and it undergoes slow denaturation in the presence of varying concentrations of GdmCl. The most interesting aspect of these studies has been the abrupt reversal of the unfolding pathway at low to moderate concentrations of the denaturant, but not at high concentration. An energetic rationale for such an unprecedented feature in protein chemistry is offered.
Department of Pharmaceutical Sciences, North Dakota State University, Fargo, North Dakota 58105, USA.
Alexander K
AK
Berg
Sumathra
S
Manokaran
Daniel
D
Eiler
Joel
J
Kooren
Sanku
S
Mallik
D K
DK
Srivastava
D000581:Amidohydrolases / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D019791:Guanidine / Q000494:pharmacology*; D007700:Kinetics; D011489:Protein Denaturation; D017510:Protein Folding
D029968:Escherichia coli Proteins; D000581:Amidohydrolases; C084507:peptide deformylase; D019791:Guanidine