Equilibrium unfolding of mutant apomyoglobins carrying substitutions of conserved nonfunctional residues with alanine

Baryshnikova, E N; Balobanov, Vitaly A.; Katina, N. S.; Melnik, Bogdan S.; Долгих, Д. А.; Semisotnov, Gennady V.; Bychkova, Valentina E.

doi:10.1134/s0026893307040139

Cited by 4 publications

(1 citation statement)

References 33 publications

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“…Based on the plasmid containing a wild-type (WT) apomyoglobin gene, plasmids possessing mutant apomyoglobin gene variants with selected substitutions were derived. Recombinant proteins were isolated and purified according to the methods described in our previous studies (Baryshnikova et al, 2005b;Baryshnikova et al, 2005a;Baryshnikova et al, 2005c;Baryshnikova et al, 2007).…”

Section: Apomyoglobinmentioning

confidence: 99%

Some useful ideas for multistate protein design: Effect of amino acid substitutions on the multistate proteins stability and the rate of protein structure formation

Majorina

Melnik

Glukhov

et al. 2022

Front. Mol. Biosci.

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The design of new protein variants is usually confined to slightly “fixing” an already existing protein, adapting it to certain conditions or to a new substrate. This is relatively easy to do if the fragment of the protein to be affected, such as the active site of the protein, is known. But what if you need to “fix” the stability of a protein or the rate of its native or intermediate state formation? Having studied a large number of protein mutant forms, we have established the effect of various amino acid substitutions on the energy landscape of the protein. As a result, we have revealed a number of patterns to help researchers identify amino acid residues that determine the folding rate and the stability of globular proteins states and design a mutant form of a protein with desired properties.

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Section: Apomyoglobinmentioning

confidence: 99%

Some useful ideas for multistate protein design: Effect of amino acid substitutions on the multistate proteins stability and the rate of protein structure formation

Majorina

Melnik

Glukhov

et al. 2022

Front. Mol. Biosci.

Self Cite

View full text Add to dashboard Cite

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sw ApoMb Amyloid Aggregation under Nondenaturing Conditions: The Role of Native Structure Stability

Katina

Balobanov

Ilyina

et al. 2017

Biophysical Journal

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Investigation of the molecular mechanisms underlying amyloid-related human diseases attracts close attention. These diseases, the number of which currently is above 40, are characterized by formation of peptide or protein aggregates containing a cross-β structure. Most of the amyloidogenesis mechanisms described so far are based on experimental studies of aggregation of short peptides, intrinsically disordered proteins, or proteins under denaturing conditions, and studies of amyloid aggregate formations by structured globular proteins under conditions close to physiological ones are still in the initial stage. We investigated the effect of amino acid substitutions on propensity of the completely helical protein sperm whale apomyoglobin (sw ApoMb) for amyloid formation from its structured state in the absence of denaturing agents. Stability and aggregation of mutated sw ApoMb were studied using circular dichroism, Fourier transform infrared spectroscopy, x-ray diffraction, native electrophoresis, and electron microscopy techniques. Here, we demonstrate that stability of the protein native state determines both protein aggregation propensity and structural peculiarities of formed aggregates. Specifically, structurally stable mutants show low aggregation propensity and moderately destabilized sw ApoMb variants form amyloids, whereas their strongly destabilized mutants form both amyloids and nonamyloid aggregates.

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Real-time tracking of protein unfolding with time-resolved x-ray solution scattering

Henry

Panman

Isaksson

et al. 2020

Structural Dynamics

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The correct folding of proteins is of paramount importance for their function, and protein misfolding is believed to be the primary cause of a wide range of diseases. Protein folding has been investigated with time-averaged methods and time-resolved spectroscopy, but observing the structural dynamics of the unfolding process in real-time is challenging. Here, we demonstrate an approach to directly reveal the structural changes in the unfolding reaction. We use nano- to millisecond time-resolved x-ray solution scattering to probe the unfolding of apomyoglobin. The unfolding reaction was triggered using a temperature jump, which was induced by a nanosecond laser pulse. We demonstrate a new strategy to interpret time-resolved x-ray solution scattering data, which evaluates ensembles of structures obtained from molecular dynamics simulations. We find that apomyoglobin passes three states when unfolding, which we characterize as native, molten globule, and unfolded. The molten globule dominates the population under the conditions investigated herein, whereas native and unfolded structures primarily contribute before the laser jump and 30 μ s after it, respectively. The molten globule retains much of the native structure but shows a dynamic pattern of inter-residue contacts. Our study demonstrates a new strategy to directly observe structural changes over the cause of the unfolding reaction, providing time- and spatially resolved atomic details of the folding mechanism of globular proteins.

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Equilibrium unfolding of mutant apomyoglobins carrying substitutions of conserved nonfunctional residues with alanine

Cited by 4 publications

References 33 publications

Some useful ideas for multistate protein design: Effect of amino acid substitutions on the multistate proteins stability and the rate of protein structure formation

Some useful ideas for multistate protein design: Effect of amino acid substitutions on the multistate proteins stability and the rate of protein structure formation

sw ApoMb Amyloid Aggregation under Nondenaturing Conditions: The Role of Native Structure Stability

Real-time tracking of protein unfolding with time-resolved x-ray solution scattering

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