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Mixture of Macromolecular Crowding Agents Has a Non-additive Effect on the Stability of Proteins

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Abstract

The folding and unfolding of proteins inside a cell take place in the presence of macromolecules of various shapes and sizes. Such crowded conditions can significantly affect folding, stability, and biophysical properties of proteins. Thus, to logically mimic the intracellular environment, the thermodynamic stability of two different proteins (lysozyme and α-lactalbumin) was investigated in the presence of mixtures of three crowding agents (ficoll 70, dextran 70, and dextran 40) at different pH values. These crowders possess different shapes and sizes. It was observed that the stabilizing effect of mixtures of crowders is more than the sum effects of the individual crowder, i.e., the stabilizing effect is non-additive in nature. Moreover, dextran 40 (in the mixture) has been found to exhibit the greatest stabilization when compared with other crowders in the mixture. In other words, the small size of the crowder has been observed to be a dominant factor in stabilization of the proteins.

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Abbreviations

GdmCl:

Guanidinium chloride

UV:

Ultra-violet

T m :

Midpoint of thermal denaturation

ΔH m :

Enthalpy change at Tm

ΔC p :

Constant-pressure heat capacity change

∆G D°:

Gibbs free energy change at 25 °C

F70:

Ficoll 70

D70:

Dextran 70

D40:

Dextran 40

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Funding

This work was supported by grant from the Science & Engineering Research Board (SERB), India (SR/FT/LS-48/2010), FIST Program (SR/FST/LSI-541/2012), and Council of Scientific and Industrial Research (CSIR), India (37(1604)/13/EMR-II). SS is thankful to Maulana Azad National Fellowship, University Grants Commission (Government of India), for providing fellowship. FA is grateful to Indian National Science Academy for the award of Senior Scientist Position.

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  1. Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, Jamia Nagar, New Delhi, 110025, India

    Sumra Shahid, Faizan Ahmad, Md. Imtaiyaz Hassan & Asimul Islam

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  1. Sumra Shahid
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  2. Faizan Ahmad
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Shahid, S., Ahmad, F., Hassan, M.I. et al. Mixture of Macromolecular Crowding Agents Has a Non-additive Effect on the Stability of Proteins. Appl Biochem Biotechnol 188, 927–941 (2019). https://doi.org/10.1007/s12010-019-02972-9

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