Thermodynamic Partitioning Model for Hydrophobic Binding of Polypeptides by GroEL

Zahn, Ralph; Axmann, Sabine E.; Rücknagel, Karl Peter; Jaeger, Ernst; Laminet, Axel A.; Plückthun, Andreas

doi:10.1006/jmbi.1994.1566

Cited by 77 publications

(57 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, it seemed likely that a collapsed, native-like structure had been formed rapidly and was recognized. Interestingly, mature p-lactamase, lacking the signal peptide, did not bind to GroEL when diluted from urea, even when the folding buffer contained sufficient denaturant to populate a known folding intermediate (Zahn & Pluckthun, 1992;Zahn et al, 1994a). At elevated temperatures (>37 "C), however, mature P-lactamase unfolded into a form that bound tightly to GroEL (Zahn & Pluckthun, 1994).…”

Section: Conformations Recognized By Groelmentioning

confidence: 99%

“…A recent, more-detailed examination of this bound intermediate, both by denaturation (monitored by Trp fluorescence) and by hydrogen-deuterium exchange (monitored by mass spectrometry), revealed considerable structure to be present, greater than that in the acid-induced molten globule conformation (Gervasoni et al, 1996). Furthermore, the observation that the apparent dissociation constant for the GroEL-P-lactamase complex decreased with increasing temperature indicated that hydrophobic interactions between p-lactamase and chaperonin were largely responsible for the binding (Zahn & Pluckthun, 1994). In another example, studies of refolding of a thermophilic lactate dehydrogenase (LDH) showed that different intermediate states could be populated upon dilution from various concentrations of guanidine hydrochloride.…”

Section: Conformations Recognized By Groelmentioning

confidence: 99%

“…Early work showed that substrate polypeptide in these complexes was exquisitely susceptible to digestion by added protease, suggesting a loosely packed, accessible structure of the bound intermediate (Martin et al, 1991;Mendoza et al, 1992a). Fluorescence studies suggested that tryptophan side chains were present in environments that were less polar than in the fully unfolded state, but more polar than when fully buried in the native state (Martin et al, 1991;Mendoza et al, 1992a;Hayer-Hart1 et al, 1994;Weissman et al, 1994;Zahn & Pluckthun, 1994). Binding of ANS to polypeptide-GroEL complexes also suggested the presence of exposed hydrophobic surface (Martin et al, 1991;Mendoza et al, 1992a;Hayer-Hart1 et al, 1994).…”

Section: Conformations Recognized By Groelmentioning

confidence: 99%

“…Several authors have suggested that the latter mechanism, thermodynamic partitioning or coupling, may be responsible for the observed unfolding of proteins by GroEL. For example, both pre-/3-lactamase (Laminet et al, 1990;Zahn et al, 1994a) and human DHFR in the absence of ligands (Viitanen et al, 1991) are unstable, relative to an inactive, less-folded state, even in the absence of GroEL. If this state exposes significant hydrophobic segments recognizable by GroEL, binding can occur to form the more-stable binary complex.…”

Section: Conformational Changes In the Substrate Proteinmentioning

confidence: 99%

“…If this state exposes significant hydrophobic segments recognizable by GroEL, binding can occur to form the more-stable binary complex. Even for an otherwise stable protein, such as mature p-lactamase, destabilization of the native state by elevated temperature may be sufficient to favor, thermodynamically, a GroEL-bound, unfolded form (Zahn & Pluckthun, 1994;Gervasoni et al, 1996). Recent experiments have examined this problem with a chemically modified form of RNAse TI that cannot complete its folding and so is trapped in a metastable state in equilibrium with a folding intermediate (Walter et al, 1996).…”

Section: Conformational Changes In the Substrate Proteinmentioning

confidence: 99%

See 4 more Smart Citations

GroEL‐Mediated protein folding

1997

View full text Add to dashboard Cite

I. Architecture of GroEL and GroES and the reaction pathwayA. Architecture of the chaperonins B. Reaction pathway of GroEL-GroES-mediated folding 3. 4. 5.Role of hydrophobicity in recognition Homologous proteins with differing recognition-differences in primary structure versus effects on folding Concluding remarksKeywords: chaperonin; GroES; Hsp60; kinetic partitioning; mechanism of action; X-ray structureThe early studies of Anfinsen and co-workers established that the 1973). Under physiologic conditions inside the cell, however, the primary structure of a protein contains all the information necesfolding process for many proteins, particularly those with multisary to direct the native secondary and tertiary fold (Anfinsen, domain structures, is prone to producing a variety of misfolded species and aggregates. Recent studies indicate that a specialized

show abstract

Section: Conformations Recognized By Groelmentioning

confidence: 99%

Section: Conformations Recognized By Groelmentioning

confidence: 99%

Section: Conformations Recognized By Groelmentioning

confidence: 99%

Section: Conformational Changes In the Substrate Proteinmentioning

confidence: 99%

Section: Conformational Changes In the Substrate Proteinmentioning

confidence: 99%

See 3 more Smart Citations

GroEL‐Mediated protein folding

1997

View full text Add to dashboard Cite

show abstract

Nuclear magnetic resonance spectroscopy with the stringent substrate rhodanese bound to the single‐ring variant SR1 of the E. coli chaperonin GroEL

et al. 2011

View full text Add to dashboard Cite

Nuclear magnetic resonance (NMR) observation of the uniformly 2 H, 15 N-labeled stringent 33-kDa substrate protein rhodanese in a productive complex with the uniformly 14 N-labeled 400 kDa single-ring version of the E. coli chaperonin GroEL, SR1, was achieved with the use of transverse relaxation-optimized spectroscopy, cross-correlated relaxation-induced polarization transfer, and cross-correlated relaxation-enhanced polarization transfer. To characterize the NMR-observable parts of the bound rhodanese, coherence buildup rates by different magnetization transfer mechanisms were measured, and effects of covalent crosslinking of the rhodanese to the apical binding surface of SR1 were investigated. The results indicate that the NMR-observable parts of the SR1-bound rhodanese are involved in intracomplex rate processes, which are not related to binding and release of the substrate protein from the SR1 binding surface. Rather, they correspond to mobility of the stably bound substrate, which thus appears to include flexibly disordered polypeptide segments devoid of long-lived secondary structures or tertiary folds, as was previously observed also with the smaller substrate human dihydrofolate reductase.

show abstract

Investigation of the folding pathway of the TEM‐1 β‐lactamase

1995

View full text Add to dashboard Cite

The TEM-1 beta-lactamase is a globular protein containing 12 proline residues. The folding mechanism of this enzyme was investigated by kinetic and equilibrium experiments with the help of fluorescence spectroscopy and circular dichroism. The equilibrium denaturation of the protein induced by guanidine hydrochloride occurs in two discrete steps, indicating the existence of a thermodynamically stable intermediate state. This state is 5.2 +/- 0.4 kcal/mol less stable than the native conformation and 5.7 +/- 0.2 kcal/mol more stable than the fully denatured protein. This intermediate state exhibits a high content of native secondary structure elements but is devoid of specific tertiary organization; its relation to the "molten globule" is discussed. Refolding kinetic experiments revealed the existence of a transient intermediate conformation between the thermodynamically stable intermediate and the native protein. This transient intermediate appears rapidly during the folding reaction. It exhibits a secondary structure content very similar to that of the native protein and has also recovered a significant amount of tertiary organisation. The final refolding step of the TEM-1 beta-lactamase, leading to the native enzyme, is dominated by two major slow kinetic phases which probably reflect a very complex process kinetically limited by proline cis/trans isomerization.

show abstract

Thermodynamic Partitioning Model for Hydrophobic Binding of Polypeptides by GroEL

Cited by 77 publications

References 17 publications

GroEL‐Mediated protein folding

GroEL‐Mediated protein folding

Nuclear magnetic resonance spectroscopy with the stringent substrate rhodanese bound to the single‐ring variant SR1 of the E. coli chaperonin GroEL

Investigation of the folding pathway of the TEM‐1 β‐lactamase

Contact Info

Product

Resources

About