Characterisation of SEQ0694 (PrsA/PrtM) of Streptococcus equi as a functional peptidyl-prolyl isomerase affecting multiple secreted protein substrates

AbstractFklB is a member of the FK506-binding proteins (FKBPs), a family that consists of five genes in Escherichia coli. Little is known about the physiological and functional role of FklB in bacterial movement. In the present study, FklB knock-out mutant ΔfklB presented an increased swarming and swimming motility and biofilm formation phenotype, suggesting that FklB is a negative regulator of these cellular processes. Complementation with Peptidyl-prolyl isomerase (PPIase)-deficient fklB gene (Y181A) revealed that the defects in biofilm formation were not restored by Y181A, indicating that PPIase activity of FklB is modulating biofilm formation in E. coli. The mean cell length of ΔfklB swarming cells was significantly smaller as compared to the wild-type BW25113. Furthermore, the mean cell length of swarming and swimming wild-type and ΔfklB cells overexpressing fklB or Y181A was considerably larger, suggesting that PPIase activity of FklB plays a role in cell elongation and/or cell division. A multi-copy suppression assay demonstrated that defects in motility and biofilm phenotype were compensated by overexpressing sets of PPIase-encoding genes. Taken together, our data represent the first report demonstrating the involvement of FklB in cellular functions of E. coli.

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Cyclophilin A Peptidyl-Prolyl Isomerase Activity Promotes Zpr1 Nuclear Export

Molecular and Cellular Biology ◽

10.1128/mcb.22.20.6993-7003.2002 ◽

2002 ◽

Vol 22 (20) ◽

pp. 6993-7003 ◽

Cited By ~ 46

Author(s):

Husam Ansari ◽

Giampaolo Greco ◽

Jeremy Luban

Keyword(s):

Nuclear Export ◽

Biological Function ◽

Zinc Finger Protein ◽

Cyclophilin A ◽

Kinetic Studies ◽

Wild Type ◽

Prolyl Isomerase ◽

Ppiase Activity ◽

Peptidyl Prolyl Isomerase ◽

Isomerase Activity

ABSTRACT The peptidyl-prolyl isomerase (PPIase) cyclophilin A (Cpr1p) is conserved from eubacteria to mammals, yet its biological function has resisted elucidation. Unable to identify a phenotype that is suggestive of Cpr1p's function in a cpr1Δ Saccharomyces cerevisiae strain, we screened for CPR1-dependent strains. In all cases, dependence was conferred by mutations in ZPR1, a gene encoding an essential zinc finger protein. CPR1 dependence was suppressed by overexpression of EF1α (a translation factor that binds Zpr1p), Cpr6p (another cyclophilin), or Fpr1p (a structurally unrelated PPIase). Suppression by a panel of cyclophilin A mutants correlated with PPIase activity, confirming the relevance of this activity in CPR1-dependent strains. In CPR1 + cells, wild-type Zpr1p was distributed equally between the nucleus and cytoplasm. In contrast, proteins encoded by CPR1-dependent alleles of ZPR1 accumulated in the nucleus, as did wild-type Zpr1p in cpr1Δ cells. Transport kinetic studies indicated that nuclear export of Zpr1p was defective in cpr1Δ cells, and rescue of this defect correlated with PPIase activity. Our results demonstrate a functional interaction between Cpr1p, Zpr1p, and EF1α, a role for Cpr1p in Zpr1p nuclear export, and a biological function for Cpr1p PPIase activity.

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Structure of an Hsp90-immunophilin complex reveals cochaperone recognition of the client-maturation state

10.1101/2021.01.21.427690 ◽

2021 ◽

Author(s):

Kanghyun Lee ◽

Aye C. Thwin ◽

Eric Tse ◽

Stephanie N. Gates ◽

Daniel R. Southworth

Keyword(s):

Prolyl Isomerase ◽

Ppiase Activity ◽

Peptidyl Prolyl Isomerase ◽

Recognition Element ◽

Middle Domain ◽

Ppiase Domain ◽

Hsp90 Chaperone ◽

Client Proteins ◽

Maturation State ◽

Tpr Domain

SummaryThe Hsp90 chaperone promotes the folding and activation of hundreds of client proteins in the cell through an ATP-dependent conformational cycle guided by distinct cochaperone regulators. The FKBP51 immunophilin binds Hsp90 with its tetratricopeptide repeat (TPR) domain and catalyzes peptidyl-prolyl isomerase (PPIase) activity during the folding of kinases, nuclear receptors and tau. Here we have determined the cryo-EM structure of the human Hsp90:FKBP51:p23 complex to 3.3 Å that, together with mutagenesis and crosslinking analysis, reveals the basis for cochaperone binding to Hsp90 during client maturation. A helix extension in the TPR functions as a key recognition element, interacting across the Hsp90 C-terminal dimer interface presented in the closed, ATP conformation. The PPIase domain is positioned along the middle domain, adjacent Hsp90 client binding sites, while a single p23 makes stabilizing interactions with the N-terminal dimer. With this architecture, FKBP51 could thereby act on specific client residues presented during Hsp90-catalyzed remodeling.

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A novel FK-506-binding-like protein that lacks peptidyl-prolyl isomerase activity is involved in intracellular infection and in vivo virulence of Burkholderia pseudomallei

Microbiology ◽

10.1099/mic.0.049163-0 ◽

2011 ◽

Vol 157 (9) ◽

pp. 2629-2638 ◽

Cited By ~ 17

Author(s):

Isobel H. Norville ◽

Katrin Breitbach ◽

Kristin Eske-Pogodda ◽

Nicholas J. Harmer ◽

Mitali Sarkar-Tyson ◽

...

Keyword(s):

Burkholderia Pseudomallei ◽

Pathogenic Bacteria ◽

Plaque Assay ◽

Bacterial Pathogen ◽

Virulence Determinants ◽

Prolyl Isomerase ◽

Ppiase Activity ◽

Fk 506 ◽

Peptidyl Prolyl Isomerase ◽

Intracellular Infection

Burkholderia pseudomallei is a facultative intracellular bacterial pathogen causing melioidosis, an often fatal infectious disease that is endemic in several tropical and subtropical areas around the world. We previously described a Ptk2 cell-based plaque assay screening system of B. pseudomallei transposon mutants that led to the identification of several novel virulence determinants. Using this approach we identified a mutant with reduced plaque formation in which the BPSL0918 gene was disrupted. BPSL0918 encodes a putative FK-506-binding protein (FKBP) representing a family of proteins that typically possess peptidyl-prolyl isomerase (PPIase) activity. A B. pseudomallei ΔBPSL0918 mutant showed a severely impaired ability to resist intracellular killing and to replicate within primary macrophages. Complementation of the mutant fully restored its ability to grow intracellularly. Moreover, B. pseudomallei ΔBPSL0918 was significantly attenuated in a murine model of infection. Structural modelling confirmed a modified FKBP fold of the BPSL0918-encoded protein but unlike virulence-associated FKBPs from other pathogenic bacteria, recombinant BPSL0918 protein did not possess PPIase activity in vitro. In accordance with this observation BPSL0918 exhibits several mutations in residues that have been proposed to mediate PPIase activity in other FKBPs. To our knowledge this B. pseudomallei FKBP represents the first example of this protein family which lacks PPIase activity but is important in intracellular infection of a bacterial pathogen.

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Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90

Bioscience Reports ◽

10.1042/bsr20150124 ◽

2015 ◽

Vol 35 (5) ◽

Cited By ~ 7

Author(s):

Elizabeth A. Blackburn ◽

Martin A. Wear ◽

Vivian Landré ◽

Vikram Narayan ◽

Jia Ning ◽

...

Keyword(s):

Heat Shock Protein 90 ◽

Prolyl Isomerase ◽

Temperature Dependent ◽

Ppiase Activity ◽

Peptidyl Prolyl Isomerase ◽

Isomerase Activity ◽

C Terminus ◽

Hsp 90 ◽

Cyclophilin 40 ◽

Tpr Domain

Binding the C-terminus of heat shock protein 90 (Hsp 90) to the tetratricopeptide repeat (TPR) domain of cyclophilin 40 (Cyp40) allosterically changes the dynamics of the cyclophilin-active site and reduces peptidyl-prolyl isomerase (PPIase) activity.

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The Peptidyl-Prolyl Isomerase Activity of SlyD Is Not Required for Maturation of Escherichia coli Hydrogenase

Journal of Bacteriology ◽

10.1128/jb.00922-07 ◽

2007 ◽

Vol 189 (21) ◽

pp. 7942-7944 ◽

Cited By ~ 20

Author(s):

Jie Wei Zhang ◽

Michael R. Leach ◽

Deborah B. Zamble

Keyword(s):

Escherichia Coli ◽

Metal Cluster ◽

The Other ◽

Prolyl Isomerase ◽

Ppiase Activity ◽

Peptidyl Prolyl Isomerase ◽

Isomerase Activity ◽

Hydrogenase Enzymes

ABSTRACT Escherichia coli SlyD, which is involved in the biosynthesis of the metal cluster in the [NiFe]-hydrogenase enzymes, exhibits several activities including that of a peptidyl-prolyl isomerase (PPIase). Mutations that result in deficient PPIase activity do not produce corresponding decreases in the other activities of SlyD in vitro or in hydrogenase production levels in vivo.

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Effect of FKBP65, a putative elastin chaperone, on the coacervation of tropoelastin in vitro

Biochemistry and Cell Biology ◽

10.1139/o10-137 ◽

2010 ◽

Vol 88 (6) ◽

pp. 917-925 ◽

Cited By ~ 10

Author(s):

Kevin L.Y. Cheung ◽

Matthew Bates ◽

Vettai S. Ananthanarayanan

Keyword(s):

Self Assembly ◽

Secretory Pathway ◽

Molar Ratio ◽

Prolyl Isomerase ◽

Ppiase Activity ◽

Peptidyl Prolyl Isomerase ◽

Maturation Stages ◽

Turbidimetric Assay

FKBP65 is a protein of the endoplasmic reticulum that is relatively abundant in elastin-producing cells and is associated with tropoelastin in the secretory pathway. To test an earlier suggestion by Davis and co-workers that FKBP65 could act as an intracellular chaperone for elastin, we obtained recombinant FKBP65 (rFKBP65) by expressing it in E. coli and examined its effect on the coacervation characteristics of chicken aorta tropoelastin (TE) using an in vitro turbidimetric assay. Our results reveal that rFKBP65 markedly promotes the initiation of coacervation of TE without significantly affecting the temperature of onset of coacervation. This effect shows saturation at a 1:2 molar ratio of TE to rFKBP65. By contrast, FKBP12, a peptidyl prolyl isomerase, has a negligible effect on TE coacervation. Moreover, the effect of rFKBP65 on TE coacervation is unaffected by the addition of rapamycin, an inhibitor of peptidyl prolyl isomerase (PPIase) activity. These observations rule out the involvement of the PPIase activity of rFKBP65 in modulating the coacervation of TE. Additional experiments using a polypeptide model of TE showed that rFKBP65, while promoting coacervation, may retard the maturation of this model polypeptide into larger aggregates. Based on these results, we suggest that FKBP65 may act as an elastin chaperone in vivo by controlling both the coacervation and the maturation stages of its self-assembly into fibrils.

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The Peptidyl-Prolyl Isomerase Motif Is Lacking in PmpA, the PrsA-Like Protein Involved in the Secretion Machinery of Lactococcus lactis

Applied and Environmental Microbiology ◽

10.1128/aem.68.8.3932-3942.2002 ◽

2002 ◽

Vol 68 (8) ◽

pp. 3932-3942 ◽

Cited By ~ 25

Author(s):

Sophie Drouault ◽

Jamila Anba ◽

Sophie Bonneau ◽

Alexander Bolotin ◽

S. Dusko Ehrlich ◽

...

Keyword(s):

Lactococcus Lactis ◽

Degradation Products ◽

Prolyl Isomerase ◽

Gene Encoding ◽

Ppiase Activity ◽

Peptidyl Prolyl Isomerase ◽

Gram Positive Bacteria ◽

The Family ◽

Almost All ◽

Cell Surface Protease

ABSTRACT The prsA-like gene from Lactococcus lactis encoding its single homologue to PrsA, an essential protein triggering the folding of secreted proteins in Bacillus subtilis, was characterized. This gene, annotated pmpA, encodes a lipoprotein of 309 residues whose expression is increased 7- to 10-fold when the source of nitrogen is limited. A slight increase in the expression of the PrsA-like protein (PLP) in L. lactis removed the degradation products previously observed with the Staphylococcus hyicus lipase used as a model secreted protein. This shows that PmpA either triggers the folding of the secreted lipase or activates its degradation by the cell surface protease HtrA. Unlike the case for B. subtilis, the inactivation of the gene encoding PmpA reduced only slightly the growth rate of L. lactis in standard conditions. However, it almost stopped its growth when the lipase was overexpressed in the presence of salt in the medium. Like PrsA of B. subtilis and PrtM of L. lactis, the L. lactis PmpA protein could thus have a foldase activity that facilitates protein secretion. These proteins belong to the third family of peptidyl-prolyl cis/trans-isomerases (PPIases) for which parvulin is the prototype. Almost all PLP from gram-positive bacteria contain a domain with the PPIase signature. An exception to this situation was found only in Streptococcaceae, the family to which L. lactis belongs. PLP from Streptococcus pneumoniae and Enterococcus faecalis possess this signature, but those of L. lactis, Streptococcus pyogenes, and Streptococcus mutans do not. However, secondary structure predictions suggest that the folding of PLP is conserved over the entire length of the proteins, including the unconserved signature region. The activity associated with the expression of PmpA in L. lactis and these genomic data show that either the PPIase motif is not necessary for PPIase activity or, more likely, PmpA foldase activity does not necessarily require PPIase activity.

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A Functional Analysis of the Cyclophilin Repertoire in the Protozoan Parasite Trypanosoma Cruzi

Biomolecules ◽

10.3390/biom8040132 ◽

2018 ◽

Vol 8 (4) ◽

pp. 132 ◽

Cited By ~ 3

Author(s):

Alina Perrone ◽

Natalia Milduberger ◽

Alicia Fuchs ◽

Patricia Bustos ◽

Jacqueline Bua

Keyword(s):

Trypanosoma Cruzi ◽

Protozoan Parasite ◽

The United States ◽

Prolyl Isomerase ◽

Ppiase Activity ◽

Peptidyl Prolyl Isomerase ◽

Isomerase Activity ◽

Evolution Of Metabolic Pathways

Trypanosoma cruzi is the etiological agent of Chagas disease. It affects eight million people worldwide and can be spread by several routes, such as vectorborne transmission in endemic areas and congenitally, and is also important in non-endemic regions such as the United States and Europe due to migration from Latin America. Cyclophilins (CyPs) are proteins with enzymatic peptidyl-prolyl isomerase activity (PPIase), essential for protein folding in vivo. Cyclosporin A (CsA) has a high binding affinity for CyPs and inhibits their PPIase activity. CsA has proved to be a parasiticidal drug on some protozoa, including T. cruzi. In this review, we describe the T. cruzi cyclophilin gene family, that comprises 15 paralogues. Among the proteins isolated by CsA-affinity chromatography, we found orthologues of mammalian CyPs. TcCyP19, as the human CyPA, is secreted to the extracellular environment by all parasite stages and could be part of a complex interplay involving the parasite and the host cell. TcCyP22, an orthologue of mitochondrial CyPD, is involved in the regulation of parasite cell death. Our findings on T. cruzi cyclophilins will allow further characterization of these processes, leading to new insights into the biology, the evolution of metabolic pathways, and novel targets for anti-T. cruzi control.

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Development of a mariner-Based Transposon and Identification of Listeria monocytogenes Determinants, Including the Peptidyl-Prolyl Isomerase PrsA2, That Contribute to Its Hemolytic Phenotype

Journal of Bacteriology ◽

10.1128/jb.00016-09 ◽

2009 ◽

Vol 191 (12) ◽

pp. 3950-3964 ◽

Cited By ~ 70

Author(s):

Jason Zemansky ◽

Benjamin C. Kline ◽

Joshua J. Woodward ◽

Jess H. Leber ◽

Hélène Marquis ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Virulence Factor ◽

Ex Vivo ◽

Physiological Role ◽

Listeriolysin O ◽

Prolyl Isomerase ◽

Peptidyl Prolyl Isomerase ◽

Growth Defects ◽

Production Activity ◽

Cell Spread

ABSTRACT Listeriolysin O (LLO) is a pore-forming toxin that mediates phagosomal escape and cell-to-cell spread of the intracellular pathogen Listeria monocytogenes. In order to identify factors that control the production, activity, or secretion of this essential virulence factor, we constructed a Himar1 mariner transposon delivery system and screened 50,000 mutants for a hypohemolytic phenotype on blood agar plates. Approximately 200 hypohemolytic mutants were identified, and the 51 most prominent mutants were screened ex vivo for intracellular growth defects. Eight mutants with a phenotype were identified, and they contained insertions in the following genes: lmo0964 (similar to yjbH), lmo1268 (clpX), lmo1401 (similar to ymdB), lmo1575 (similar to ytqI), lmo1695 (mprF), lmo1821 (similar to prpC), lmo2219 (prsA2), and lmo2460 (similar to cggR). Some of these genes are involved in previously unexplored areas of research with L. monocytogenes: the genes yjbH and clpX regulate the disulfide stress response in Bacillus subtilis, and the prpC phosphatase has been implicated in virulence in other gram-positive pathogens. Here we demonstrate that prsA2, an extracytoplasmic peptidyl-prolyl cis/trans isomerase, is critical for virulence and contributes to the folding of LLO and to the activity of another virulence factor, the broad-range phospholipase C (PC-PLC). Furthermore, although it has been shown that prsA2 expression is linked to PrfA, the master virulence transcription factor in L. monocytogenes pathogenesis, we demonstrate that prsA2 is not directly controlled by PrfA. Finally, we show that PrsA2 is involved in flagellum-based motility, indicating that this factor likely serves a broad physiological role.

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