Testing polyols' compatibility with Gibbs energy of stabilization of proteins under conditions in which they behave as compatible osmolytes.


Abstract

It is generally believed that compatible osmolytes stabilize proteins by shifting the denaturation equilibrium, native state <--> denatured state toward the left. We show here that if osmolytes are compatible with the functional activity of the protein at a given pH and temperature, they should not significantly perturb this denaturation equilibrium under the same experimental conditions. This conclusion was reached from the measurements of the activity parameters (K(m) and k(cat)) and guanidinium chloride-induced denaturations of lysozyme and ribonuclease-A in the presence of five polyols (sorbitol, glycerol, mannitol, xylitol and adonitol) at pH 7.0 and 25 degrees C. Study holds ProTherm entries: 20618, 20619, 20620, 20621, 20622, 20623, 20624, 20625, 20626, 20627, 20628, 20629, 20630, 20631, 20632, 20633, 20634, 20635, 20636, 20637, 20638, 20639, 20640, 20641, 20642, 20643, 20644, 20645, 20646, 20647, 20648, 20649, 20650, 20651, 20652, 20653, 20654, 20655, 20656, 20657, 20658, 20659, 20660, 20661, 20662 Extra Details: Protein stability; Guanidinium chloride denaturation; Enzyme activity; Polyol osmolytes; Lysozyme; Ribonuclease-A

Submission Details

ID: tXmZkqQt

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:53 p.m.

Version: 1

Publication Details
Haque I;Singh R;Ahmad F;Moosavi-Movahedi AA,FEBS Lett. (2005) Testing polyols' compatibility with Gibbs energy of stabilization of proteins under conditions in which they behave as compatible osmolytes. PMID:15990095
Additional Information

Number of data points 90
Proteins Lysozyme C ; Ribonuclease pancreatic ; Ribonuclease pancreatic ; Lysozyme C
Unique complexes 2
Assays/Quantities/Protocols Experimental Assay: Cm details:Additives Mannitol (1.00 M) ; Experimental Assay: dG_H2O details:Additives Mannitol (1.00 M) ; Experimental Assay: Cm details:Additives Adonitol (1.00 M) ; Experimental Assay: dG_H2O details:Additives Adonitol (1.00 M) ; Experimental Assay: Cm details:Additives Xylitol (1.00 M) ; Experimental Assay: dG_H2O details:Additives Xylitol (1.00 M) ; Experimental Assay: Cm details:Additives Xylitol (0.75 M) ; Experimental Assay: dG_H2O details:Additives Xylitol (0.75 M) ; Experimental Assay: Cm details:Additives Glycerol (4.10 M) ; Experimental Assay: dG_H2O details:Additives Glycerol (4.10 M) ; Experimental Assay: Cm details:Additives Sorbitol (1.65 M) ; Experimental Assay: dG_H2O details:Additives Sorbitol (1.65 M) ; Experimental Assay: Cm details:Additives Mannitol (0.75 M) ; Experimental Assay: dG_H2O details:Additives Mannitol (0.75 M) ; Experimental Assay: Cm details:Additives Mannitol (0.50 M) ; Experimental Assay: dG_H2O details:Additives Mannitol (0.50 M) ; Experimental Assay: Cm details:Additives Mannitol (0.25 M) ; Experimental Assay: dG_H2O details:Additives Mannitol (0.25 M) ; Experimental Assay: Cm details:Additives Adonitol (0.75 M) ; Experimental Assay: dG_H2O details:Additives Adonitol (0.75 M) ; Experimental Assay: Cm details:Additives Adonitol (0.50 M) ; Experimental Assay: dG_H2O details:Additives Adonitol (0.50 M) ; Experimental Assay: Cm details:Additives Adonitol (0.25 M) ; Experimental Assay: dG_H2O details:Additives Adonitol (0.25 M) ; Experimental Assay: Cm details:Additives xylitol (0.75 M) ; Experimental Assay: dG_H2O details:Additives xylitol (0.75 M) ; Experimental Assay: Cm details:Additives Xylitol (0.50 M) ; Experimental Assay: dG_H2O details:Additives Xylitol (0.50 M) ; Experimental Assay: Cm details:Additives Xylitol (0.25 M) ; Experimental Assay: dG_H2O details:Additives Xylitol (0.25 M) ; Experimental Assay: Cm details:Additives Glycerol (2.74 M) ; Experimental Assay: dG_H2O details:Additives Glycerol (2.74 M) ; Experimental Assay: Cm details:Additives Glycerol (1.37 M) ; Experimental Assay: dG_H2O details:Additives Glycerol (1.37 M) ; Experimental Assay: Cm details:Additives Sorbitol (1.10 M), ; Experimental Assay: dG_H2O details:Additives Sorbitol (1.10 M), ; Experimental Assay: Cm details:Additives Sorbitol (0.55 M) ; Experimental Assay: dG_H2O details:Additives Sorbitol (0.55 M) ; Experimental Assay: Cm details:Additives ; Experimental Assay: dG_H2O details:Additives
Libraries Mutations for sequence KVFGRCELAAAMKRHGLDNYRGYSLGNWVCAAKFESNFNTQATNRNTDGSTDYGILQINSRWWCNDGRTPGSRNLCNIPCSALLSSDITASVNCAKKIVSDGNGMNAWVAWRNRCKGTDVQAWIRGCRL ; Mutations for sequence KETAAAKFERQHMDSSTSAASSSNYCNQMMKSRNLTKDRCKPVNTFVHESLADVQAVCSQKNVACKNGQTNCYQSYSTMSITDCRETGSSKYPNCAYKTTQANKHIIVACEGNPYVPVHFDASV
Sequence Assay Result Units