title: Selective inactivation of parvulin-like peptidyl-prolyl cis/trans isomerases by juglone.
abstract: In contrast to FK506 binding proteins and cyclophilins, the parvulin family of peptidyl-prolyl cis/trans isomerases (PPIases; E.C. 5.2.1.8) cannot be inhibited by either FK506 or cyclosporin A. We have found that juglone, 5-hydroxy-1,4-naphthoquinone, irreversibly inhibits the enzymatic activity of several parvulins, like the E. coli parvulin, the yeast Ess1/Ptf1, and human Pin1, in a specific manner, thus allowing selective inactivation of these enzymes in the presence of other PPIases. The mode of action was studied by analyzing the inactivation kinetics and the nature of products of the reaction of E. coli parvulin and its Cys69Ala variant with juglone. For all parvulins investigated, complete inactivation was obtained by a slow process that is characterized by pseudo-first-order rate constants in the range of 5.3 x 10(-)4 to 4. 5 x 10(-)3 s-1. The inactivated parvulin contains two juglone molecules that are covalently bound to the side chains of Cys41 and Cys69 because of a Michael addition of the thiol groups to juglone. Redox reactions did not contribute to the inactivation process. Because thiol group modification was shown to proceed 5-fold faster than the rate of enzyme inactivation, it was considered as a necessary but not sufficient condition for inactivation. When measured by far-UV circular dichroism (CD), the rate of structural alterations following thiol group modification parallels exactly the rate of inactivation. Thus, partial unfolding of the active site of the parvulins was thought to be the cause of the deterioration of PPIase activity.
authors.affiliation: Max-Planck Research Unit, Enzymology of Protein Folding, Halle/Saale, Germany.
authors.forename: L
authors.initials: L
authors.lastname: Hennig
authors.forename: C
authors.initials: C
authors.lastname: Christner
authors.forename: M
authors.initials: M
authors.lastname: Kipping
authors.forename: B
authors.initials: B
authors.lastname: Schelbert
authors.forename: K P
authors.initials: KP
authors.lastname: Rücknagel
authors.forename: S
authors.initials: S
authors.lastname: Grabley
authors.forename: G
authors.initials: G
authors.lastname: Küllertz
authors.forename: G
authors.initials: G
authors.lastname: Fischer
mesh_terms: D000591:Amino Acid Isomerases / Q000037:antagonists & inhibitors; D019943:Amino Acid Substitution / Q000235:genetics; D002942:Circular Dichroism; D003545:Cysteine / Q000235:genetics; D010450:Endopeptidases; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins; D005656:Fungal Proteins / Q000037:antagonists & inhibitors; D005978:Glutathione / Q000737:chemistry; D006801:Humans; D006868:Hydrolysis; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D009285:Naphthoquinones / Q000494:pharmacology*; D019696:Peptidylprolyl Isomerase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D013053:Spectrophotometry; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014157:Transcription Factors / Q000037:antagonists & inhibitors
chemical_list: D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D005656:Fungal Proteins; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D009285:Naphthoquinones; D013438:Sulfhydryl Compounds; D014157:Transcription Factors; D010450:Endopeptidases; D000591:Amino Acid Isomerases; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase; C000606851:parvA protein, E coli; D005978:Glutathione; D003545:Cysteine; C005134:juglone