Engineered disulfide bonds as probes of the folding pathway of barnase: Increasing the stability of proteins against the rate of denaturation

Disulfide bonds and calcium ions contribute significantly to the stability of the hemocyanin from the mollusc Rapana thomasiana grosse (gastropod). An extremely powerful protective effect of Ca2+ at a concentration of 100 mᴍ (100% protection) against the destructive effect of reductants like dithiothreitol was observed. This is important for the practical application of molluscan hemocyanins in experimental biochemistry, immunology and medicine. The reduction of the disulfide bonds in the Rapana hemocyanin leads to a 20% decrease of the α-helical structure. The S-S bonds contribute significantly to the free energy of stabilization in water increasing ⊿GD H2O by 6.9 kJ mol-1. The data are related to the X-ray model of the Rapana hemocyanin functional unit RtH2e. The results of this study can be of common validity for related respiratory proteins because the cysteine residues are conserved in all sequences of molluscan hemocyanins published so far.

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Context-dependent effects of proline residues on the stability and folding pathway of ubiquitin

European Journal of Biochemistry ◽

10.1111/j.1432-1033.2004.04392.x ◽

2004 ◽

Vol 271 (22) ◽

pp. 4474-4484 ◽

Cited By ~ 16

Author(s):

Maria D. Crespo ◽

Geoffrey W. Platt ◽

Roger Bofill ◽

Mark S. Searle

Keyword(s):

Folding Pathway ◽

Context Dependent ◽

The Stability

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The Differential Impact of Disulfide Bonds and N-Linked Glycosylation on the Stability and Function of CD14

Journal of Biological Chemistry ◽

10.1074/jbc.m707640200 ◽

2007 ◽

Vol 283 (6) ◽

pp. 3376-3384 ◽

Cited By ~ 19

Author(s):

Jianmin Meng ◽

Peggy Parroche ◽

Douglas T. Golenbock ◽

C. James McKnight

Keyword(s):

Disulfide Bonds ◽

Differential Impact ◽

The Stability ◽

And Function

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COMPUTER SIMULATIONS TO INVESTIGATE STABILITY AND STRUCTURAL PROPERTIES OF PEPTIDE AMPHIPHILE NANOFIBERS

Journal of Theoretical and Computational Chemistry ◽

10.1142/s021963360700326x ◽

2007 ◽

Vol 06 (03) ◽

pp. 621-630

Author(s):

RUO-YU CHEN ◽

LING-YING WU ◽

JUN-MIN LIAO ◽

CHENG-LUNG CHEN

Keyword(s):

Molecular Dynamics ◽

Disulfide Bonds ◽

Md Simulation ◽

Intermolecular Hydrogen Bond ◽

Intermolecular Distance ◽

Peptide Amphiphile ◽

Hydrogen Bond Interactions ◽

Self Assembling ◽

Polar Groups ◽

The Stability

Molecular mechanics (MM) method followed by molecular dynamics (MD) simulation was carried out to investigate the stability of an aggregate formed by self-assembling of peptide amphiphile (PA) molecules. The MM + MD simulation confirms that the cylindrical shaped aggregate is very stable. The analysis showed that the remarkable stability of the aggregate was partly due to various intermolecular hydrogen-bond interactions between polar groups of PA molecules. The hydrophobic alkyl tails of PA molecules are packed loosely inside the interior of the aggregates. The packing of alkyl tails contribute further stability of the PA aggregate. Our simulations reproduce qualitatively experimental observations and support the fact that PA molecules are self-assembled within closed intermolecular distance to favor the forming of disulfide bonds.

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Rational Design of Protein Stability: Effect of (2S,4R)-4-Fluoroproline on the Stability and Folding Pathway of Ubiquitin

PLoS ONE ◽

10.1371/journal.pone.0019425 ◽

2011 ◽

Vol 6 (5) ◽

pp. e19425 ◽

Cited By ~ 29

Author(s):

Maria D. Crespo ◽

Marina Rubini

Keyword(s):

Protein Stability ◽

Rational Design ◽

Folding Pathway ◽

The Stability ◽

Stability Effect

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Structural Analysis of Avian Encephalomyelitis Virus Polyprotein for Development of Multi Epitopes Vaccine Using Immunoinformatics Approach

Journal of Pure and Applied Microbiology ◽

10.22207/jpam.15.1.20 ◽

2021 ◽

Vol 15 (1) ◽

pp. 262-278

Author(s):

Fatima Khalid Elhassan ◽

Yassir A. Almofti ◽

Khoubieb Ali Abd-elrahman ◽

Mashair AA Nouri ◽

Elsideeq EM Eltilib

Keyword(s):

T Cells ◽

Disulfide Bonds ◽

Tertiary Structure ◽

High Mobility ◽

Dynamics Simulation ◽

High Morbidity ◽

E Coli ◽

Encephalomyelitis Virus ◽

The Stability ◽

Chimeric Vaccine

Avian Encephalomyelitis (AE) is the disease caused by avian encephalomyelitis virus (AEV). The disease mainly affects young birds nervous system worldwide causing high morbidity and variable mortality rate in chicks and noticed egg dropping and hatchability in mature hens. Vaccination is the only way to control AEV infection since there is no treatment yet to the avian encephalomyelitis. This study aimed to use immunoinformatics approaches to predict multi epitopes vaccine from the AEV polyprotein that could elicit both B and T cells. The vaccine construct comprises 482 amino acids obtained from epitopes predicted against B and T cells by IEDB server, adjuvant, linkers and 6-His-tag. The chimeric vaccine was potentially antigenic and nonallergic and demonstrated thermostability and hydrophilicity in protparam server. The solubility of the vaccine was measured in comparison to E. coli proteins. The stability was also assessed by disulﬁde bonds engineering to reduce the high mobility regions in the designed vaccine. Furthermore molecular dynamics simulation further strengthen stability of the predicted vaccine. Tertiary structure of the vaccine construct after prediction, refinement was used for molecular docking with chicken alleles BF2*2101 and BF2*0401 and the docking process demonstrated favourable binding energy score of -337.47 kcal/mol and -326.87 kcal/mol, respectively. Molecular cloning demonstrated the potential clonability of the chimeric vaccine in pET28a(+) vector. This could guarantee the efficient translation and expression of the vaccine within suitable expression vector.

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Dynamic transitions in the translocated phloem filament protein

Functional Plant Biology ◽

10.1071/pp99161 ◽

2000 ◽

Vol 27 (9) ◽

pp. 733 ◽

Cited By ~ 5

Author(s):

Kirsten Leineweber ◽

Alexander Schulz ◽

Gary A. Thompson

Keyword(s):

Disulfide Bonds ◽

Assimilate Transport ◽

Phloem Sap ◽

Folding Intermediates ◽

P Protein ◽

Molecular Masses ◽

P Proteins ◽

The Stability ◽

Filament Protein ◽

Higher Sensitivity

This paper originates from a presentation at the International Conference on Assimilate Transport and Partitioning, Newcastle, NSW, August 1999 Recent evidence suggests that the P-proteins of Cucurbita maxima exist in at least two structural states: large polymers that are immobilized in individual sieve elements and small polymers or individual subunits that are translocated over long distances. We investigated variation in the structure of the phloem filament protein (phloem protein 1 or PP1) to determine the translocated form of the protein and its relationship to the polymerized state. It was demonstrated that the stability, folding state and assembly of the phloem filament protein rely on distinct intramolecular disulfide bonds. Acid trapping experiments combined with intergeneric grafts revealed that the phloem filament protein is translocated as an 88 kDa globular protein. By altering the pH of the collection buffer (pH 2–10), four individual conformational isoforms of PP1 with molecular masses of 81, 83, 85 and 88 kDa were consistently observed. The 81 kDa isoform represents the totally reduced phloem filament protein, the 83 and 85 kDa isoforms folding intermediates, and the 88 kDa its native soluble translocated form. The 83 and 85 kDa folding intermediates are susceptible to aggregation causing the gelation and formation of P-protein filaments in oxidized phloem sap. In contrast to the 88 kDa globular transport form, the 81, 83 and 85 kDa isoforms possibly exhibit lower stability, and therefore a higher sensitivity to proteolytic digestion.

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