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Employing in vitro analysis to test the potency of methylglyoxal in inducing the formation of amyloid-like aggregates of caprine brain cystatin

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Abstract

Thiol protease inhibitors (cystatins) are implicated in various disease states from cancer to neurodegenerative conditions and immune responses. Cystatins have high amyloidogenic propensity and they are prone to form fibrillar aggregates leading to amyloidosis. Particularly challenging examples of such disorders occur in type 2 diabetes, Alzheimer’s and Parkinson’s diseases. The aim of the present study is to find an interaction between the compound methylglyoxal (MG) which is particularly elevated in type 2 diabetes with caprine brain cystatin (CBC). Results have shown that elevated concentration of MG forms amyloid aggregates of CBC. This was achieved by allowing slow growth in a solution containing moderate to high concentrations of MG. When analysed with microscopy, the protein aggregate present in the sample after incubation consisted of extended filaments with ordered structures. This fibrillar material possesses extensive β-sheet structure as revealed by far-UV CD and IR spectroscopy. Furthermore, the fibrils exhibit increased Thioflavin T fluorescence.

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Abbreviations

MG:

Methylglyoxal

CBC:

Caprine brain cystatin

AGEs:

Advanced glycation end products

AD:

Alzheimer disease

PD:

Parkinson’s disease

mM:

Millimolar

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Acknowledgments

The work was supported by the project of Scientific Council of Industrial Research (CSIR) India; vide no. 37(1520)/12/EMR-II. We would like to thank All India Institute of Medical sciences, New Delhi (AIIMS) for TEM analysis, facilities available at AMU Aligarh are highly acknowledged.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, 202002, UP, India

    Waseem Feeroze Bhat, Sheraz Ahmad Bhat, Peerzada Shariq Shaheen Khaki & Bilqees Bano

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  1. Waseem Feeroze Bhat
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  2. Sheraz Ahmad Bhat
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Correspondence to Bilqees Bano.

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Bhat, W.F., Bhat, S.A., Khaki, P.S.S. et al. Employing in vitro analysis to test the potency of methylglyoxal in inducing the formation of amyloid-like aggregates of caprine brain cystatin. Amino Acids 47, 135–146 (2015). https://doi.org/10.1007/s00726-014-1848-2

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