title: Analysis of surface cavity in serpin family reveals potential binding sites for chemical chaperone to reduce polymerization.
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.
authors.affiliation: Protein Conformation and Enzymology Lab, Department of Biosciences, Jamia Millia Islamia University, New-Delhi 110025, India.
authors.forename: Poonam
authors.initials: P
authors.lastname: Singh
authors.forename: Mohammad Sazzad
authors.initials: MS
authors.lastname: Khan
authors.forename: Asma
authors.initials: A
authors.lastname: Naseem
authors.forename: Mohamad Aman
authors.initials: MA
authors.lastname: Jairajpuri
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D000991:Antithrombins / Q000737:chemistry; D001665:Binding Sites; D002241:Carbohydrates / Q000737:chemistry; D058105:Polymerization; D011480:Protease Inhibitors; D011485:Protein Binding; D011487:Protein Conformation; D015842:Serine Proteinase Inhibitors / Q000737:chemistry; D015843:Serpins / Q000737:chemistry*
chemical_list: D000596:Amino Acids; D000991:Antithrombins; D002241:Carbohydrates; D011480:Protease Inhibitors; D015842:Serine Proteinase Inhibitors; D015843:Serpins