Truncated GroEL monomer has the ability to promote folding of rhodanese without GroES and ATP

Makino, Yoshihide; Taguchi, Hideki; Yoshida, Masasuke

doi:10.1016/0014-5793(93)80838-l

Cited by 34 publications

(21 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, it has been observed that folding of hen lysozyme can be promoted by GroEL alone, in the absence of GroES and ATP (20). Similar results have been demonstrated for barnase (21) and for rhodanese (22).…”

supporting

confidence: 88%

Accelerated folding in the weak hydrophobic environment of a chaperonin cavity: Creation of an alternate fast folding pathway

Jewett

Baumketner

Shea

2004

Proc. Natl. Acad. Sci. U.S.A.

111

158

View full text Add to dashboard Cite

Recent experiments suggest that the folding of certain proteins can take place entirely within a chaperonin-like cavity. These substrate proteins experience folding rate enhancements without undergoing multiple rounds of ATP-induced binding and release from the chaperonin. Rather, they undergo only a single binding event, followed by sequestration into the chaperonin cage. The present work uses molecular dynamics simulations to investigate the folding of a highly frustrated protein within this chaperonin cavity. The chaperonin interior is modeled by a sphere with a lining of tunable degree of hydrophobicity. We demonstrate that a moderately hydrophobic environment, similar to the interior of the GroEL cavity upon complexion with ATP and GroES, is sufficient to accelerate the folding of a frustrated protein by more than an order of magnitude. Our simulations support a mechanism by which the moderately hydrophobic chaperonin environment provides an alternate pathway to the native state through a transiently bound intermediate state.

show abstract

supporting

confidence: 88%

Accelerated folding in the weak hydrophobic environment of a chaperonin cavity: Creation of an alternate fast folding pathway

Jewett

Baumketner

Shea

2004

Proc. Natl. Acad. Sci. U.S.A.

111

158

View full text Add to dashboard Cite

show abstract

“…One model of GroEL action [39] assumes the possibility of interaction of free monomers with non-folded proteins, followed by the formation of the 7-mers and 14-mers. A chaperone activity of GroEL monomers different from that of the 14-mers has recently been discovered [41]. It is a question for future studies whether the dynamic Mg-ATP-dependent transitions between the monomeric and 14-meric forms can play a particular role in the functioning of the chaperonins.…”

Section: Mg-atp (Adp)mentioning

confidence: 99%

In vitro dissociation and self‐assembly of three chaperonin 60s: the role of ATP

Lissin

1995

FEBS Letters

View full text Add to dashboard Cite

A comparative study has investigated the in vitro dissociation and self-assembly of chaperonin 60 14-mers isolated from E. coli (GroEL), yeast mitochondria and pea chloroplasts. In all cases Mg ~+ inhibits, and low temperature stimulates, the urea-induced dissociation. ATP or ADP in the presence of Mg 2÷ enhance the dissociation of the chaperonins. Re-assembly of the 14-mers from their monomers shows different efficiencies between the three proteins. In all cases, however, self-assembly is stimulated by Mg-adenine nucleotides. Surprisingly, effective self-assembly of GroEL is promoted by 20% glycerol in the absence of ATP. The role of Mg-adenine nucleotides in the dissociation and assembly of the chaperonins is discussed.

show abstract

“…The protein GroEL was purified by modifications of the procedure included in Ref. 13. After ammonium sulfate fractionation, the protein was purified by means of three chromatographic steps: DEAE-Sephacel, gel filtration through Sepharose CL-4B, and affinity chromatography through red agarose (14).…”

Section: Methodsmentioning

confidence: 99%

Recognition of Protein Substrates by Protein-disulfide Isomerase

Cheung

Churchich

1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

Refolding of partially folded mitochondrial malate dehydrogenase (mMDH) is assisted by protein-disulfide isomerase (PDI).It is shown that the fluorescence probe is covalently attached to a SH residue located in the b domain. Based on the fluorescence measurements of native and derivatized PDI, it is suggested that recognition of the unfolded substrate involves conformational changes propagated to several domains of PDI. Protein-disulfide isomerase (PDI)1 is a multifunctional enzyme that both catalyzes the formation of disulfide bonds (1, 2) and acts as a subunit of prolyl-4-hydroxylase (3). PDI has been proposed to function as a molecular chaperone by binding to unfolded protein species, thereby preventing aggregation and misfolding (4, 5). Despite these interesting studies, the situation is complicated because PDI, unlike other chaperones, catalyzes disulfide bond formation and reduction.Several laboratories have attempted to show the site of binding of small molecular weight polypeptides that compete with misfolded protein substrates. Mutated PDI with the carboxyl terminus deleted shows neither peptide binding nor chaperone activity in assisting the refolding of denatured D-glyceraldehyde-3-phosphate dehydrogenase (6). On the other hand, it has been reported that deletion of the carboxyl-terminal domain (C domain) has no inhibitory effect on the assembly of recombinant prolyl-4-hydroxylase (7). Other investigators have reported that small molecular weight peptides bind to the bЈ domain of PDI (8).The possibility exists that more than one site in the structure of PDI is involved in recognition and refolding of protein substrates. The binding and hydrolysis of ATP by PDI has been reported by Guthapfel et al. (9); strikingly, the ATPase reaction is stimulated in the presence of denatured polypeptides, whereas the disulfide oxidation of PDI is not influenced by ATP. However, the functional role played by ATP hydrolysis during the refolding of denatured proteins has not been investigated in detail.The aim of the present work is 2-fold: first, to study regions of the primary structure of PDI involved in recognition of unfolded protein substrates and, second, to investigate whether the free energy of hydrolysis of ATP is required for unfolding of misfolded protein substrates. EXPERIMENTAL PROCEDURESPurification of Proteins-PDI was purified by the method described in Ref. 10 with small modifications. Fresh porcine livers (600 g) were homogenized in 0.1 M phosphate (pH 7.5) containing 1% Triton X-100 and 5 mM EDTA. After centrifugation, the supernatant was treated with ammonium sulfate, and the fractions obtained between 55-85% saturation were suspended in 25 mM citrate buffer (pH 5.3), dialyzed against the same buffer (buffer A), applied to CM-Sephadex C-50 column and eluted with the same buffer. Fractions displaying PDI activity were pooled, dialyzed against 20 mM sodium phosphate (pH 6.3) (buffer B), and applied to a DEAE-Sepharose fast flow column, which was eluted using a linear gradient of 0 -0.7 M NaCl in buffer...

show abstract

Truncated GroEL monomer has the ability to promote folding of rhodanese without GroES and ATP

Cited by 34 publications

References 27 publications

Accelerated folding in the weak hydrophobic environment of a chaperonin cavity: Creation of an alternate fast folding pathway

Accelerated folding in the weak hydrophobic environment of a chaperonin cavity: Creation of an alternate fast folding pathway

In vitro dissociation and self‐assembly of three chaperonin 60s: the role of ATP

Recognition of Protein Substrates by Protein-disulfide Isomerase

Contact Info

Product

Resources

About