Dependence of the anti-chaperone activity of protein disulphide isomerase on its chaperone activity

Song, Jiu-li; Quan, Hui; Wang, Chih-chen

doi:10.1042/bj3280841

Cited by 30 publications

(21 citation statements)

References 27 publications

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“…Thus the anti-chaperone activity of PDI at low concentrations resulted from noncovalent cross-linking rather than the cross-linking of the disulfide bonds. These observations were quite consistent with those in previous studies (11,20,21).…”

Section: Pdi Exhibits Both Anti-chaperone and Chaperone Activity Insupporting

confidence: 83%

“…These results suggested that the formation of disulfide bonds between PDI and CK contributed to the suppression of CK aggregation during refolding but was not the major driving force. DISCUSSION PDI has been demonstrated to act as a molecular chaperone independent of its isomerase activity (21)(22)(23)(24)(25), and it can exploit the synergy between these functions, especially when disulfide reshuffling is needed in the refolding of the substrates (3,26). Our results showed that WT PDI at high concentrations could act as a protector against aggregation but acted as an inhibitor of reactivation during CK refolding.…”

Section: Pdi Exhibits Both Anti-chaperone and Chaperone Activity Inmentioning

confidence: 70%

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Catalysis of Creatine Kinase Refolding by Protein Disulfide Isomerase Involves Disulfide Cross-link and Dimer to Tetramer Switch

Zhao

Xie

et al. 2005

Journal of Biological Chemistry

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Protein disulfide isomerase (PDI) functions as an isomerase to catalyze thiol:disulfide exchange, as a chaperone to assist protein folding, and as a subunit of prolyl-4-hydroxylase and microsomal triglyceride transfer protein. At a lower concentration of 0.2 M, PDI facilitated the aggregation of unfolded rabbit muscle creatine kinase (CK) and exhibited anti-chaperone activity, which was shown to be mainly due to the hydrophobic interactions between PDI and CK and was independent of the cross-linking of disulfide bonds. At concentrations above 1 M, PDI acted as a protector against aggregation but an inhibitor of reactivation during CK refolding. The inhibition effect of PDI on CK reactivation was further characterized as due to the formation of PDI-CK complexes through intermolecular disulfide bonds, a process involving Cys-36 and Cys-295 of PDI. Two disulfide-linked complexes containing both PDI and CK were obtained, and the large, soluble aggregates around 400 kDa were composed of 1 molecule of tetrameric PDI and 2 molecules of inactive intermediate dimeric CK, whereas the smaller one, around 200 kDa, was formed by 1 dimeric PDI and 1 dimeric CK. To our knowledge this is the first study revealing that PDI could switch its conformation from dimer to tetramer in its functions as a foldase. According to the observations in this research and our previous study of the folding pathways of CK, a working model was proposed for the molecular mechanism of CK refolding catalyzed by PDI.In eukaryotes, all the nascent outer membrane and secreted proteins are synthesized and fold to their native structures in the lumen of endoplasmic reticulum. The specialized compartment provides a favorable environment for native disulfide bond formation, which is often the rate-limiting step in the folding of many proteins. Endoplasmic reticulum is abundant in molecular chaperones and folding catalysts, among which the most abundant and efficient catalyst of disulfide bond formation is protein disulfide isomerase (PDI, 1 EC 5.3.4.1) (1-3). PDI is a multidomain and multifunction homodimer. It comprises four thioredoxin-like domains, a, b, bЈ, and aЈ, plus a linker region between bЈ and aЈ and a C-terminal acid extension (2, 4, 5). Each of the two catalytic domains, a and aЈ, contains a WCGHCK motif, responsible for the isomerase activity (6). In vitro studies showed that the bЈ domain provides the principle peptide binding site, whereas the other domains should also be involved when the protein substrates have a substantial secondary structure (7, 8). The latest study (9) demonstrated the existence of a ligand binding site in the bЈ domain, which was a small hydrophobic pocket defined as Much effort has been devoted to investigating the catalytic mechanism of PDI since the first report published more than 40 years ago (10). As an enzyme, the specific binding to its substrate is a basic property of PDI, and the nature of the interaction between PDI and its substrates plays a crucial role in its activity of accelerating steps in protein fol...

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Section: Pdi Exhibits Both Anti-chaperone and Chaperone Activity Insupporting

confidence: 83%

Section: Pdi Exhibits Both Anti-chaperone and Chaperone Activity Inmentioning

confidence: 70%

Catalysis of Creatine Kinase Refolding by Protein Disulfide Isomerase Involves Disulfide Cross-link and Dimer to Tetramer Switch

Zhao

Xie

et al. 2005

Journal of Biological Chemistry

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“…This was also confirmed by recent work of Dai and Wang who showed that truncation of the last 50 amino acids of PDI, comprising part of the C-terminal ␣ helix of the a domain, led to inhibition of the chaperone and peptide binding activities (24).…”

Section: The Inactive Full-length F449r Mutant Regains Peptide Bindinsupporting

confidence: 70%

Mutations That Destabilize the a′ Domain of Human Protein-disulfide Isomerase Indirectly Affect Peptide Binding

Klappa¹,

Koivunen²,

Pirneskoski³

et al. 2000

Journal of Biological Chemistry

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Protein-disulfide isomerase (PDI) is a catalyst of folding of disulfide-bonded proteins and also a multifunctional polypeptide that acts as the ␤-subunit in the prolyl 4-hydroxylase ␣ 2 ␤ 2 -tetramer (P4H) and the microsomal triglyceride transfer protein ␣␤-dimer. The principal peptide-binding site of PDI is located in the b domain, but all domains contribute to the binding of misfolded proteins. Mutations in the C-terminal part of the a domain have significant effects on the assembly of the P4H tetramer and other functions of PDI. In this study we have addressed the question of whether these mutations in the C-terminal part of the a domain, which affect P4H assembly, also affect peptide binding to PDI. We observed a strong correlation between P4H assembly competence and peptide binding; mutants of PDI that failed to form a functional P4H tetramer were also inactive in peptide binding. However, there was also a correlation between inactivity in these assays and indicators of conformational disruption, such as protease sensitivity. Peptide binding activity could be restored in inactive, protease-sensitive mutants by selective proteolytic removal of the mutated a domain. Hence we propose that structural changes in the a domain indirectly affect peptide binding to the b domain. Protein-disulfide isomerase (PDI),1 an abundant protein of the eukaryotic endoplasmic reticulum, is a catalyst of disulfide bond formation and rearrangement in the course of protein folding (for review see Ref. 1). A molecular interpretation of PDI activity is made difficult by the fact that PDI is multifunctional in the cell. In addition, it has a wide range of actions in vitro and consists of multiple domains. Furthermore, there is as yet no high resolution structure of full-length PDI. Current efforts are focused on analyzing the domain structure of PDI and in establishing the roles of specific domains in specific functional activities.It is now clear (2, 3) that PDI has a structural organization based on duplicated sequence modules (Fig. 1). The full-length protein is constructed of four structural domains with homologous thioredoxin folds plus a C-terminal acidic extension. The homologous a and a sequence modules contain the active site motif -WCGHC-and show significant sequence identity to thioredoxin; a high resolution NMR analysis of the recombinant a domain confirms that it has the thioredoxin fold. The homologous b and b modules do not show significant sequence similarity to the a domain, but NMR analysis has revealed that the b domain also exhibits the thioredoxin fold (2, 4). Analysis of the properties of individual domains as catalysts of simple thiol:disulfide oxidoreduction and of more complex protein folding linked to disulfide isomerization shows that the a and a domains function effectively as simple thiol:disulfide oxidoreductases but that the remaining domains are required for full activity in catalyzing protein folding associated with the formation of native disulfide bonds (5). No specific function has yet been ascribed...

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“…The nature of this interaction is most likely hydrophobic since it was greatly diminished by the presence of EG. EG increases the hydrophobicity of the solution, thereby decreasing the hydrophobic interaction between folding intermediate and trigger factor~Li & Zhou, 1997;Song et al, 1997!. Rapid dilution of the GdnHCl denatured GAPDH to nondenaturing conditions allowed at least three noncovalent processes to occur: intramolecular organization of peptide segments~folding!, intermolecular assembly of hydrophobic surfaces~aggregation!, and formation of the trigger factor0 GAPDH complex. The result of this competition may be determined by either kinetic or thermodynamic factors.…”

Section: Discussionmentioning

confidence: 99%

Assisted folding of D‐glyceraldehyde‐3‐phosphate dehydrogenase by trigger factor

Huang

Zhou

et al. 2000

Protein Science

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The Escherichia coli trigger factor is a peptidyl-prolyl cis-trans isomerase that catalyzes proline-limited protein folding extremely well. Here, refolding of d-glyceraldehyde-3-phosphate dehydrogenase~GAPDH! in the presence of trigger factor was investigated. The regain of activity of GAPDH was markedly increased by trigger factor after either longor short-term denaturation, and detectable aggregation of GAPDH intermediates was prevented. In both cases, time courses of refolding of GAPDH were decelerated by trigger factor. The reactivation yield of GAPDH showed a slow down-turn when molar ratios of trigger factor to GAPDH were above 5, due to tight binding between trigger factor and GAPDH intermediates. Such inactive bound GAPDH could be partially rescued from trigger factor by addition of reduced aLA as competitor, by further diluting the refolding mixture, or by disrupting hydrophobic interactions in the complexes. A model for trigger factor assisted refolding of GAPDH is proposed. We also suggest that assisted refolding of GAPDH is due mainly to the chaperone function of trigger factor.

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Dependence of the anti-chaperone activity of protein disulphide isomerase on its chaperone activity

Cited by 30 publications

References 27 publications

Catalysis of Creatine Kinase Refolding by Protein Disulfide Isomerase Involves Disulfide Cross-link and Dimer to Tetramer Switch

Catalysis of Creatine Kinase Refolding by Protein Disulfide Isomerase Involves Disulfide Cross-link and Dimer to Tetramer Switch

Mutations That Destabilize the a′ Domain of Human Protein-disulfide Isomerase Indirectly Affect Peptide Binding

Assisted folding of D‐glyceraldehyde‐3‐phosphate dehydrogenase by trigger factor

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