title: Guanidine-HCl dependent structural unfolding of M-crystallin: fluctuating native state like topologies and intermolecular association.
abstract: Numerous experimental techniques and computational studies, proposed in recent times, have revolutionized the understanding of protein-folding paradigm. The complete understanding of protein folding and intermediates are of medical relevance, as the aggregation of misfolding proteins underlies various diseases, including some neurodegenerative disorders. Here, we describe the unfolding of M-crystallin, a βγ-crystallin homologue protein from archaea, from its native state to its denatured state using multidimensional NMR and other biophysical techniques. The protein, which was earlier characterized to be a predominantly β-sheet protein in its native state, shows different structural propensities (α and β), under different denaturing conditions. In 2 M GdmCl, the protein starts showing two distinct sets of peaks, with one arising from a partially unfolded state and the other from a completely folded state. The native secondary structural elements start disappearing as the denaturant concentration approaches 4 M. Subsequently, the protein is completely unfolded when the denaturant concentration is 6 M. The (15)N relaxation data (T(1)/T(2)), heteronuclear (1)H-(15)N Overhauser effects (nOes), NOESY data, and other biophysical data taken together indicate that the protein shows a consistent, gradual change in its structural and motional preferences with increasing GdmCl concentration.
authors.affiliation: Department of Chemical Sciences, Tata Institute of Fundamental Research, Colaba, Mumbai, India.
authors.forename: Ravi Pratap
authors.initials: RP
authors.lastname: Barnwal
authors.forename: Geetika
authors.initials: G
authors.lastname: Agarwal
authors.forename: Kandala V R
authors.initials: KV
authors.lastname: Chary
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism; D003459:Crystallins / Q000737:chemistry* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D019791:Guanidine / Q000494:pharmacology*; D008958:Models, Molecular; D011489:Protein Denaturation / Q000187:drug effects*; D017433:Protein Structure, Secondary / Q000187:drug effects; D013696:Temperature
chemical_list: D019843:Archaeal Proteins; D003459:Crystallins; D019791:Guanidine