‘Molten‐globule“ state accumulates in carbonic anhydrase folding

Долгих, Д. А.; Kolomiets, A.P.; Bolotina, I. A.; Ptitsyn, Oleg B.

doi:10.1016/0014-5793(84)80020-4

Cited by 179 publications

(131 citation statements)

References 16 publications

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“…Originally the molten globule was defined as a "compact globule with native-like secondary structure and with slowly fluctuating tertiary structure" (7), where the side chains were believed to be "liquid-like" (6,37). The experimental data supporting this claim was the absence of a near-UV CD signal and sharp NMR signals from the aromatic groups, indicating that the aromatic side chains undergo conformational averaging (6,37,38). The data presented here demonstrate that the hydrophobic core is structured in the molten globule NCBD.…”

Section: Discussionmentioning

confidence: 99%

Conformational selection in the molten globule state of the nuclear coactivator binding domain of CBP

Kjærgaard

Teilum

Poulsen

2010

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

153

242

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Native molten globules are the most folded kind of intrinsically disordered proteins. Little is known about the mechanism by which native molten globules bind to their cognate ligands to form fully folded complexes. The nuclear coactivator binding domain (NCBD) of CREB binding protein is particularly interesting in this respect as structural studies of its complexes have shown that NCBD folds into two remarkably different states depending on the ligand being ACTR or IRF-3. The ligand-free state of NCBD was characterized in order to understand the mechanism of folding upon ligand binding. Biophysical studies show that despite the molten globule nature of the domain, it contains a small cooperatively folded core. By NMR spectroscopy, we have demonstrated that the folded core of NCBD has a well ordered conformer with specific side chain packing. This conformer resembles the structure of the NCBD in complex with the protein ligand, ACTR, suggesting that ACTR binds to prefolded NCBD molecules from the ensemble of interconverting structures.CREB binding protein | folding upon binding | NMR spectroscopy T he protein structure-function paradigm has dominated structural and molecular biology over the past five decades. This paradigm has been challenged by the discovery of functional but intrinsically disordered proteins (IDPs) (1). IDPs are abundant in higher organisms and are necessary for many crucial biological functions such as cell cycle regulation, signal transduction, and regulation of transcription (2, 3). Structural studies of IDPs have shown that despite the high degree of disorder, these proteins are far from randomly structured and form transient, yet specific, secondary and tertiary structural elements (4, 5). The success of the structure-function paradigm in explaining the function of folded proteins suggests that the functions of the IDPs may also be understood by studying the transient structure of the disordered state.The molten globule was originally discovered as a partially folded state under mildly denaturing conditions and was shown to accumulate as an intermediate during protein folding reactions (6, 7). With the emergence of the IDPs, proteins were discovered where the molten globule is the functionally active state. Native molten globules are the most compact conformational state considered as IDPs (4). Molten globules have native-like secondary structure, but are believed to lack the well-defined tertiary interactions found in folded proteins. Most high resolution NMR studies of the molten globule state have focused on the secondary structure of the backbone as this is more readily accessible from secondary chemical shifts (8). The degree of side chain packing, however, is less well understood. Molten globules are believed to have dynamic hydrophobic cores due to the lack of signal in the near-UV CD spectrum and the broadening of NMR signals of aromatic groups (6, 7). Recent studies have shown that the side chains in the α-lactalbumin molten globule exchange between several well defined confor...

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Section: Discussionmentioning

confidence: 99%

Conformational selection in the molten globule state of the nuclear coactivator binding domain of CBP

Kjærgaard

Teilum

Poulsen

2010

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

153

242

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“…The most interesting examples of such 'partly unfolded' states are the 'molten globule' states which have been observed for a number of proteins [25-331 at low pH, high temperatures, etc. These states can be slightly [25][26][27]341 or more 1351 expanded as compared with the native state but all of them are much more compact than the statistical coil and possess native-like secondary structure. However they lack rigid tertiary structure as indicated by circular dichroism of aromatic groups (fluorescence and circular dichroism) and NMR spectra.…”

Section: The Molten Globule State As the Denatured State Required Formentioning

confidence: 99%

The ‘molten globule’ state is involved in the translocation of proteins across membranes?

1988

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Strong evidence exists that the translocation of proteins across a variety of membranes involves ti non-native or denatured conformational states. On the other hand a compact state having secondary but not rigid tertiary structure and called the 'molten globule' state has been identified as being stable under mild denaturing conditions. A similar state has been shown to accumulate on the folding pathway of globular proteins. These states are compact though sufficiently expanded to include water, and they are internally mobile. It is proposed that these molten globule states may be suitable candidates for protein translocation across biological membranes.

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“…CD studies by manual mixing have shown [6,7] that the secondary structure of carbonic anhydrase is restored in less than 100s. CD measurements with the stopped-flow technique have not yet been performed.…”

Section: Febs Letters November 1987mentioning

confidence: 99%

“…In particular, it has been shown that refolding of carbonic anhydrase B proceeds through an intermediate state [4][5][6][7] which has been identified as a 'molten globule' state [7]. However, the refolding kinetics was studied at time intervals >100 s, therefore the formation of early kinetic intermediates could not be followed.…”

Section: Introductionmentioning

confidence: 99%

Sequential mechanism of refolding of carbonic anhydrase B

et al. 1987

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The kinetics of refolding of bovine carbonic anhydrase B was studied by a variety of methods over a wide range of times (from milliseconds to hours). It has been shown that protein refolding proceeds through three stages. At the first stage (tl/2~0.03 s) hydrophobic clusters and a compact state of the chain are formed. At the second stage (tl,'2 ~ 140 s) hydrophobic clusters are desolvated and the rigid native-like hydrophobic core is formed. At the third stage (tl/2 ~ 600 s) the native active protein is formed.

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‘Molten‐globule“ state accumulates in carbonic anhydrase folding

Cited by 179 publications

References 16 publications

Conformational selection in the molten globule state of the nuclear coactivator binding domain of CBP

Conformational selection in the molten globule state of the nuclear coactivator binding domain of CBP

The ‘molten globule’ state is involved in the translocation of proteins across membranes?

Sequential mechanism of refolding of carbonic anhydrase B

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