title: NMR insights into folding and self-association of Plasmodium falciparum P2.
abstract: The eukaryotic 60S-ribosomal stalk is composed of acidic ribosomal proteins (P1 and P2) and neutral protein P0, which are thought to be associated as a pentameric structure, [2P1, 2P2, P0]. Plasmodium falciparum P2 (PfP2) appears to play additional non-ribosomal functions associated with its tendency for homo-oligomerization. Recombinant bacterially expressed PfP2 protein also undergoes self-association, as shown by SDS-PAGE analysis and light scattering studies. Secondary structure prediction algorithms predict the native PfP2 protein to be largely helical and this is corroborated by circular dichroism investigation. The (1)H-(15)N HSQC spectrum of native P2 showed only 43 cross peaks compared to the expected 138. The observed peaks were found to belong to the C-terminal region, suggesting that this segment is flexible and solvent exposed. In 9 M urea denaturing conditions the chain exhibited mostly non-native β structural propensity. (15)N Relaxation data for the denatured state indicated substantial variation in ms-µs time scale motion along the chain. Average area buried upon folding (AABUF) calculations on the monomer enabled identification of hydrophobic patches along the sequence. Interestingly, the segments of slower motion in the denatured state coincided with these hydrophobic patches, suggesting that in the denatured state the monomeric chain undergoes transient hydrophobic collapse. The implications of these results for the folding mechanism and self-association of PfP2 are discussed.
authors.affiliation: Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai, India.
authors.forename: Pushpa
authors.initials: P
authors.lastname: Mishra
authors.forename: Sudipta
authors.initials: S
authors.lastname: Das
authors.forename: Lata
authors.initials: L
authors.lastname: Panicker
authors.forename: Madhusoodan V
authors.initials: MV
authors.lastname: Hosur
authors.forename: Shobhona
authors.initials: S
authors.lastname: Sharma
authors.forename: Ramakrishna V
authors.initials: RV
authors.lastname: Hosur
mesh_terms: D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010963:Plasmodium falciparum / Q000378:metabolism*; D017510:Protein Folding; D015800:Protozoan Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins