Unfolding of multimeric proteins in presence of denaturants. A case study of helianthinin from Helianthus annuus L.
Helianthinin (11S), a multisubunit protein from Sunflower Seeds (Helianthus annuus L.) dissociates to its monomer (2S) through a trimeric (7S) intermediate as a function of guanidine hydrochloride (GuHCl) and guanidine thiocyanate (GuHSCN) concentration. Measurements of viscosity, velocity sedimentation patterns and spectroscopic parameters of the protein in presence of these denaturants both at equilibrium and as a function of time clearly suggest that the dissociation, unfolding and aggregation of this multimeric protein occur sequentially. The unfolding of the protein in guanidinium salts has two transitions with the first transition occurring between native to intermediate and the second transition occurring between intermediate state and unfolded state. The midpoint concentration for the major transition is 1.75 M for GuHCl and 0.8 M for GuHSCN. Evaluation of this data suggests that during the process of denaturation the simultaneous unfolding of acidic and basic subunits of the protein takes place. At intermediate concentrations of denaturant namely 1.6 M of GuHCl or 1.1 M of GuHSCN the aggregation of the protein was found to be maximum. The results suggests the possibility of a mechanism for the dissociation, denaturation and unfolding of multimeric proteins in presence of the chemical denaturants.
Suryaprakash
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P
Central Food Technological Research Institute, Department of Protein Chemistry and Technology, Mysore, India.
Prakash
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V
D055516:2S Albumins, Plant; D052179:Antigens, Plant; D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry; D006146:Guanidines / Q000737:chemistry; D006368:Helianthus / Q000737:chemistry*; D007700:Kinetics; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D011489:Protein Denaturation; D017510:Protein Folding; D055314:Seed Storage Proteins; D012639:Seeds / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D013861:Thiocyanates / Q000737:chemistry; D014461:Ultracentrifugation; D014783:Viscosity
D055516:2S Albumins, Plant; D052179:Antigens, Plant; D006146:Guanidines; D010940:Plant Proteins; D055314:Seed Storage Proteins; D013861:Thiocyanates; C067568:methionine-rich 2S protein, Helianthus; C054436:guanidine thiocyanate; D019791:Guanidine