Erratum to “Inhibition of botulinum neurotoxins interchain disulfide bond reduction prevents the peripheral neuroparalysis of botulism” [Biochem. Pharmacol. 98 (2015) 522–530]

Tropomyosin (Tpm) is a coiled-coil actin-binding dimer protein that participates in the regulation of muscle contraction. Both Tpm chains contain Cys190 residues which are normally in the reduced state, but form an interchain disulfide bond in failing heart. Changes in structural and functional properties of Tpm and its complexes with actin upon disulfide cross-linking were studied using various experimental methods. To understand the molecular mechanism underlying these changes and to reveal the possible mechanism of the involvement of the cross-linking in heart failure, molecular dynamics (MD) simulations of the middle part of Tpm were performed in cross-linked and reduced states. The cross-linking increased bending stiffness of Tpm assessed from MD trajectories at 27 °C in agreement with previous experimental observations. However, at 40 °C, the cross-linking caused a decrease in Tpm stiffness and a significant reduction in the number of main chain hydrogen bonds in the vicinity of residues 133 and 134. These data are in line with observations showing enhanced thermal unfolding of the least stable part of Tpm at 30–40 °C and accelerated trypsin cleavage at residue 133 at 40 °C (but not at 27 °C) upon cross-linking. These results allow us to speculate about the possible mechanism of involvement of Tpm cross-linking to heart failure pathogenesis.

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Cytotoxic effects of ricin without an interchain disulfide bond: genetic modification and chemical crosslinking studies

Biochimica et Biophysica Acta (BBA) - General Subjects ◽

10.1016/0304-4165(94)00166-u ◽

1995 ◽

Vol 1243 (3) ◽

pp. 399-406 ◽

Cited By ~ 19

Author(s):

D. Mohanraj ◽

S. Ramakrishnan

Keyword(s):

Disulfide Bond ◽

Genetic Modification ◽

Chemical Crosslinking ◽

Cytotoxic Effects ◽

Interchain Disulfide Bond

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Effect of immunoglobulin G (IgG) interchain disulfide bond cleavage on efficacy of intravenous immunoglobulin for immune thrombocytopenic purpura (ITP)

Clinical & Experimental Immunology ◽

10.1111/j.1365-2249.2010.04255.x ◽

2010 ◽

Vol 162 (3) ◽

pp. 415-424 ◽

Cited By ~ 15

Author(s):

Y. Machino ◽

H. Ohta ◽

E. Suzuki ◽

S. Higurashi ◽

T. Tezuka ◽

...

Keyword(s):

Intravenous Immunoglobulin ◽

Disulfide Bond ◽

Immunoglobulin G ◽

Immune Thrombocytopenic Purpura ◽

Bond Cleavage ◽

Thrombocytopenic Purpura ◽

Interchain Disulfide Bond

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Quantitative Risk assessment of product disulfide bond reduction in a recombinant protein manufacturing

10.22541/au.162456077.74648055/v1 ◽

2021 ◽

Author(s):

Yi Qing cui ◽

rujie meng ◽

yutong Li

Keyword(s):

Risk Assessment ◽

Cell Culture ◽

Recombinant Protein ◽

Disulfide Bond ◽

Culture Fluid ◽

Culture Conditions ◽

Quantitative Risk Assessment ◽

Prevention Measures ◽

Recombinant Molecule ◽

Disulfide Bond Reduction

Disulfide bond reduction occurred now and then during a recombinant protein manufacturing. Their reduction often led in the loss of batches. Various prevention measures were developed. Their implementation in a manufacturing inevitably increases its operation complexity and even might impact its product qualities and process performances. It is highly desirable to apply a reduction measure only if necessary and to apply no strong measure if a moderate one works. The reduction occurrence is determined both by the reduction sensitivity of a recombinant molecule and by the reduction power of harvested cell culture fluids. The reduction power of harvested cell culture fluids varies largely and depends on cells, lysis level, growth stage, and culture conditions. The method of the quantitative risk assessment of disulfide bond reduction was reported here. The quantitative assessment is realized by comparing the reduction sensitivity of a recombinant molecule with the reduction power of its harvested cell culture fluid.

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