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Analysis of surface cavity in serpin family reveals potential binding sites for chemical chaperone to reduce polymerization

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Abstract

Serpin constitute about 10% of blood protein and are associated with mutations that results in aberrant intermolecular linkages which leads to polymer formation. Studies with short peptides have shown promise in depolymerization of serpins however a reactive center loop based peptide also makes the serpin inactive. A chemical chaperone based approach is a better option in terms of maintaining activity and retarding polymerization but not much is known about its binding and mechanism. Specific target for chemical chaperones and its effectiveness across many serpin is not known. We did an analysis of serpin cavity using CASTp and show that cavities are distributed throughout the molecule where the largest cavities are generally present in areas of major conformational change like shutter region, helix D and helix F. An analysis of different conformational states of serpins showed that this large cavity undergoes increase in size in latent and cleaved states as compared to native state. We targeted serpins with a variety of carbohydrate, methylamine and amino acid based chemical chaperones and selected those that have highest binding energy across different serpins to assess their ability to bind large cavities. The results show that carbohydrate based chemical chaperone like sorbitol, sucrose, arabitol and trehalose and amino acid based chaperones like dopamine, phenylalanine, arginine and glutamic acid are the most effective in binding serpins. Most of these chemical chaperone interacted with residues in the shutter region and the helix D arm at the C-terminal which are part of the largest cavities. We selected the carbohydrate based chemical chaperone with best binding energies and did experimental study under the condition that induce polymerization and show that indeed they were able to retard polymer formation with moderate effect on inhibition rates. However a fluorometric study with native antithrombin showed that chemical chaperone may effect the conformation of the proteins. Our study shows that chemical chaperones have the best binding affinities for the cavities around shutter region and helix D and that a cavity targeting based approach seems to be a better option for retarding polymerization in serpins, but a thorough analysis of its effect on folding, inhibition and cofactor binding is required.

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Abbreviations

Serpin:

Serine protease inhibitors

ATIII:

Antithrombin

HCFII:

Heparin cofactor II

PAI:

Plasminogen activator inhibitor

PC1:

Protein C1-inhibitor

ADT-Vina:

Autodock tools vina

CASTp:

Computed atlas of surface topography of proteins

RCL:

Reactive center loop

TMAO:

Trimethylamine N-oxide

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Acknowledgments

This research was supported by grants from Department of Biotechnology and University Grant Commission, Government of India. PS is supported by a grant from Rajiv Gandhi National Fellowship. AN is supported by an Innovation in Science Pursuit for Inspired Research (INSPIRE) fellowship from Department of Science and Technology, Government of India. SK is supported by a fellowship from University Grant Commission.

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Authors and Affiliations

  1. Protein Conformation and Enzymology Lab, Department of Biosciences, Jamia Millia Islamia University, New-Delhi, 110025, India

    Poonam Singh, Mohammad Sazzad Khan, Asma Naseem & Mohamad Aman Jairajpuri

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  1. Poonam Singh
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  2. Mohammad Sazzad Khan
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  4. Mohamad Aman Jairajpuri
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Correspondence to Mohamad Aman Jairajpuri.

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Singh, P., Khan, M.S., Naseem, A. et al. Analysis of surface cavity in serpin family reveals potential binding sites for chemical chaperone to reduce polymerization. J Mol Model 18, 1143–1151 (2012). https://doi.org/10.1007/s00894-011-1110-8

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  • DOI: https://doi.org/10.1007/s00894-011-1110-8

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