Sequential mechanism of refolding of carbonic anhydrase B

Semisotnov, Gennady V.; Rodionova, N. A.; Kutyshenko, V. P.; Ebert, Bernd; Blanck, Jfirgen; Ptitsyn, Oleg B.

doi:10.1016/0014-5793(87)80412-x

Cited by 217 publications

(205 citation statements)

References 8 publications

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“…As a further test for the possible presence of partially folded states, we examined AMB by using the dye ANS. This dye has long been utilized to probe for exposure of hydrophobic regions present in partially folded states (20), and a recent study on the formation of amyloid fibrils from acidic fibroblast growth factor demonstrates that this spectroscopic probe can be used under high-temperature conditions (21). As expected, HMB does not bind ANS at 65°C or room temperature, consistent with the absence of stable of stable hydrophobic clusters (data not shown).…”

Section: Amyloid Fibril Formation Does Not Correlate With the Presencmentioning

confidence: 52%

Myoglobin forms amyloid fibrils by association of unfolded polypeptide segments

Fändrich¹,

Forge²,

Buder³

et al. 2003

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

267

238

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Observations that ␤-sheet proteins form amyloid fibrils under at least partially denaturing conditions has raised questions as to whether these fibrils assemble by docking of preformed ␤-structure or by association of unfolded polypeptide segments. By using ␣-helical protein apomyoglobin, we show that the ease of fibril assembly correlates with the extent of denaturation. By contrast, monomeric ␤-sheet intermediates could not be observed under the conditions of fibril formation. These data suggest that amyloid fibril formation from apomyoglobin depends on disordered polypeptide segments and conditions that are selectively unfavorable to folding. However, it is inevitable that such conditions often stabilize protein folding intermediates.

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Section: Amyloid Fibril Formation Does Not Correlate With the Presencmentioning

confidence: 52%

Myoglobin forms amyloid fibrils by association of unfolded polypeptide segments

Fändrich¹,

Forge²,

Buder³

et al. 2003

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

267

238

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“…The fluorescence from ANS is amplified upon its binding to hydrophobic surfaces or clusters that are characteristics of partially structured proteins (28,29). We observed a small amplification of the ANS fluorescence with both the native (Fig.…”

Section: Resultsmentioning

confidence: 69%

Specific collapse followed by slow hydrogen-bond formation of β-sheet in the folding of single-chain monellin

Kimura

Uzawa

Ishimori

et al. 2005

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

124

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Characterization of the conformational landscapes for proteins with different secondary structures is important in elucidating the mechanism of protein folding. The folding trajectory of singlechain monellin composed of a five-stranded ␤-sheet and a helix was investigated by using a pH-jump from the alkaline unfolded to native state. The kinetic changes in the secondary structures and in the overall size and shape were measured by circular dichroism spectroscopy and small-angle x-ray scattering, respectively. The formation of the tertiary structure was monitored by intrinsic and extrinsic fluorescence. A significant collapse was observed within 300 s after the pH-jump, leading to the intermediate with a small amount of secondary and tertiary structures but with an overall oblate shape. Subsequently, the stepwise formation of secondary and tertiary structures was detected. The current observation was consistent with the theoretical prediction that a more significant collapse precedes the formation of secondary structures in the folding of ␤-sheet proteins than that of helical proteins [Shea, J. E., Onuchic, J. N. & Brooks, C. L., III (2002) Proc. Natl. Acad. Sci. USA 99, 16064 -16068]. Furthermore, it was implied that the initial collapse was promoted by the formation of some specific structural elements, such as tight turns, to form the oblate shape.energy landscape ͉ protein folding ͉ submillisecond dynamics ͉ x-ray scattering C haracterization of the folding dynamics for proteins and their free energy landscapes offers a key to understanding protein folding phenomena (1, 2). The coarse-grained approximation of intraprotein interactions (a ''funneled'' landscape) has been proposed (3, 4) and has succeeded in explaining the structures of proteins in the folding transition state (5-8). However, the molecular basis of the funneled landscape, which is realized by a complex interplay of various intraprotein interactions and polypeptide dynamics, is largely unknown. For example, although the hydrophobic interaction and hydrogen bond (H-bond) are the major stabilizing interactions for the specific folded conformations of proteins, their contribution to the energy landscape, including the unfolded conformations, is controversial (9). Furthermore, the relationship between protein folds and the characteristics of landscapes has not been well investigated. Therefore, the observation of the entire folding process of proteins with various folds is required to characterize the free energy landscapes.We previously observed the folding dynamics of ␣-helical proteins, cytochrome c (cyt c) and apomyoglobin (apoMb), based on helical content and compactness, which mainly reflect the formation of the H-bonds and hydrophobic interactions, respectively (10-12). The characterized landscapes of these proteins demonstrated the cooperative acquisition of the helical structure and compactness after the initial collapse and suggested that the hydrophobic environment created by the collapse facilitates the subsequent conformational search. ...

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“…IC) has a sharp maximum at 1.45 M Gdm-HCI. This shows that BCAB strongly binds ANS under these conditions, which is a specific test for lVlG [4,6,7,17].…”

Section: Resultsmentioning

confidence: 98%

“…It is almost as compact as the native state (N), has a pronounced secondary structure and differs from N mainly by the absence of ~ight packing of side chains in the protein core and by a substantial increase of fluctuations [1][2][3][4]. The molten globule state (MN) accumulates during the renaturation of globular proteins from the fully unfolded state (U) [1][2][3][4][5][6][7][8] and therefore may play a universal role in protein folding [7]. It has been also suggested [9] and shown experimentally that the molten globule is trapped by Gro-EL chaperons ( [10] and unpublished data of G.V.…”

Section: Introductionmentioning

confidence: 99%

‘All‐or‐none’ mechanism of the molten globule unfolding

et al. 1992

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The Gdm-HCl-induced unfolding of bovine carbonic anhydrase B and S. at~ret~s fl-lactamase was studied at 4°C by a variety of methods. With the use of FPLC it has been shown that within the transition from the molten globule to the unfolded state the distribution function of molecular dimensions is bimodal. This means that equilibrium intermediates between the molten globule and the unfolded states are about, i.¢. the molten globule unfolding follows the "all-or-none' mechanism.

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Sequential mechanism of refolding of carbonic anhydrase B

Cited by 217 publications

References 8 publications

Myoglobin forms amyloid fibrils by association of unfolded polypeptide segments

Myoglobin forms amyloid fibrils by association of unfolded polypeptide segments

Specific collapse followed by slow hydrogen-bond formation of β-sheet in the folding of single-chain monellin

‘All‐or‐none’ mechanism of the molten globule unfolding

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