The peptide backbone plays a dominant role in protein stabilization by naturally occurring osmolytes.
Transfer free energy measurements of amino acids from water to the osmolytes, sucrose and sarcosine, were made as a function of osmolyte concentration. From these data, transfer free energies of the amino acid side chains were obtained, and the transfer free energy of the peptide backbone was determined from solubility measurements of diketopiperazine (DKP). Using static accessible surface evaluations of the native and unfolded states of ribonuclease A, solvent exposed side chain and peptide backbone areas were multiplied by their transfer free energies and summed in order to evaluate the transfer free energy of the native and unfolded states of the protein from water to the osmolyte solutions. The results reproduced the main features of the free energy profile determined for denaturation of proteins in the presence of osmolytes. The side chains were found collectively to favor exposure to the osmolyte in comparison to exposure in water, and in this sense the side chains favor protein unfolding. The major factor which opposes and overrides the side chain preference for denaturation and results in the stabilization of proteins observed in osmolytes is the highly unfavorable exposure of polypeptide backbone on unfolding. Except for urea and guanidine hydrochloride solutions, it is shown that all organic solvents (e.g., dioxane, ethanol, ethylene glycol) and solutes (osmolytes) for which transfer free energy measurements have been determined exhibit unfavorable transfer free energy of the peptide backbone.(ABSTRACT TRUNCATED AT 250 WORDS)
Liu
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Y
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch at Galveston 77555-1052, USA.
Bolen
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DW
D009994:Osmolar Concentration; D010455:Peptides / Q000737:chemistry*; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D012521:Sarcosine / Q000737:chemistry*; D013395:Sucrose / Q000737:chemistry*; D013816:Thermodynamics
D010455:Peptides; D013395:Sucrose; D012259:Ribonuclease, Pancreatic; D012521:Sarcosine