4.7 Article

Conformational behavior of alpha-2-macroglobulin: Aggregation and inhibition induced by TFE

Journal

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume 104, Issue -, Pages 539-546

Publisher

ELSEVIER
DOI: 10.1016/j.ijbiomac.2017.06.026

Keywords

Aggregation; Alpha-2-macroglobulin; Proteinase inhibitor; TFE; Protein folding

Funding

  1. Department of Science and Technology (Fund for the Improvement of Science and Technology Infrastructure in Higher Educational Institutions)
  2. University Grants Commission (Special Assistance Program), Government of India
  3. UGC-BSR (Basic Scientific Research)

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Alpha-2-macroglobulin (alpha M-2), a pan-proteinase inhibitor, inhibits a variety of endogenous and exogenous proteinases and constitutes an important part of body's innate defense system. In the present study, we explored how trifluoroethanol (TFE) may modulate the structure, antiproteinase activity and aggregation of alpha M-2. TFE was sequentially added over a range of 0-20% (v/v) and the effects induced were studied by activity assay, intrinsic fluorescence, ANS fluorescence, circular dichroism, turbidity assay, Rayleigh scattering measurement and ThT fluorescence measurement. Decrease in activity and increase in fluorescence intensity of alpha M-2 upon addition of TFE shows structural deviation from the native structure and suggests aggregation of protein upon solvent addition. Increase in turbidity and Rayleigh scattering of modified alpha M-2 confirms the formation of aggregates. Insignificant ThT fluorescence intensity of TFE treated alpha M-2 is indicative of amorphous or non-amyloid aggregation. Further, circular dichroism results indicate the changes in secondary structure of native alpha M-2 as negative ellipticity decreased on addition of the polar solvent to the inhibitor. The turbidometric analysis, Rayleigh scattering, ThT fluorescence intensity of modified alpha M-2 suggests that the protein might be driven towards non-amyloid or amorphous aggregation. Our studies provide important mechanistic insight how alpha M-2 undergoes conformational and functional changes when exposed to TFE. (C) 2017 Elsevier B.V. All rights reserved.

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