title: Generation of a highly active folding enzyme by combining a parvulin-type prolyl isomerase from SurA with an unrelated chaperone domain.
abstract: Parvulins are small prolyl isomerases and serve as catalytic domains of folding enzymes. SurA (survival protein A) from the periplasm of Escherichia coli consists of an inactive (Par1) and an active (Par2) parvulin domain as well as a chaperone domain. In the absence of the chaperone domain, the folding activity of Par2 is virtually abolished. We created a chimeric protein by inserting the chaperone domain of SlyD, an unrelated folding enzyme from the FKBP family, into a loop of the isolated Par2 domain of SurA. This increased its folding activity 450-fold to a value higher than the activity of SurA, in which Par2 is linked with its natural chaperone domain. In the presence of both the natural and the foreign chaperone domain, the folding activity of Par2 was 1500-fold increased. Related and unrelated chaperone domains thus are similarly efficient in enhancing the folding activity of the prolyl isomerase Par2. A sequence analysis of various chaperone domains suggests that clusters of exposed methionine residues in mobile chain regions might be important for a generic interaction with unfolded protein chains. This binding is highly dynamic to allow frequent transfer of folding protein chains between chaperone and catalytic domains.
authors.affiliation: Laboratorium für Biochemie und Bayreuther Zentrum für Molekulare Biowissenschaften, Universität Bayreuth, D-95440 Bayreuth, Germany.
authors.forename: Anne-Juliane
authors.initials: AJ
authors.lastname: Geitner
authors.forename: Edina
authors.initials: E
authors.lastname: Varga
authors.forename: Marc
authors.initials: M
authors.lastname: Wehmer
authors.forename: Franz X
authors.initials: FX
authors.lastname: Schmid
mesh_terms: D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D004795:Enzyme Stability; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry*; D008969:Molecular Sequence Data; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D054730:Protein Interaction Domains and Motifs; D058767:Protein Unfolding; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D016415:Sequence Alignment
chemical_list: D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D011993:Recombinant Fusion Proteins; C490900:SurA protein, E coli; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase; C000606851:parvA protein, E coli
keywords: 12-kDa human FK506 binding protein; 5-(((acetylamino)ethyl)amino)naphthalene-1-sulfonic acid; AEDANS; Abz; FKBP12; GdmCl; IF; RCM-T1; RNase T1; SlyD; SurA; aminobenzoyl; artificial enzyme; domain insertion; folding catalysis; guanidinium chloride; insert-in-flap; pNA; para-nitroanilide; protein folding; ribonuclease T1; survival protein A; the reduced and carboxymethylated form of the S54G/P55N variant of ribonuclease T1