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

Study Summary

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 KETAAAKFERQHMDSSTSAASSSNYCNQMMKSRNLTKDRCKPVNTFVHESLADVQAVCSQKNVACKNGQTNCYQSYSTMSITDCRETGSSKYPNCAYKTTQANKHIIVACEGNPYVPVHFDASV ; Mutations for sequence KVFGRCELAAAMKRHGLDNYRGYSLGNWVCAAKFESNFNTQATNRNTDGSTDYGILQINSRWWCNDGRTPGSRNLCNIPCSALLSSDITASVNCAKKIVSDGNGMNAWVAWRNRCKGTDVQAWIRGCRL

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
2IHL 1993-06-29T00:00:00+0000 1.4 LYSOZYME (E.C.3.2.1.17) (JAPANESE QUAIL)
6QMN 2019-02-07T00:00:00+0000 2.31 Crystal structure of a Ribonuclease A-Onconase chimera
1FBI 1995-01-19T00:00:00+0000 3.0 CRYSTAL STRUCTURE OF A CROSS-REACTION COMPLEX BETWEEN FAB F9.13.7 AND GUINEA-FOWL LYSOZYME
1GHL 1993-05-04T00:00:00+0000 2.1 THE THREE-DIMENSIONAL STRUCTURE OF PHEASANT AND GUINEA-FOWL EGG LYSOZYMES
1HHL 1993-05-04T00:00:00+0000 1.9 THE THREE-DIMENSIONAL STRUCTURE OF PHEASANT AND GUINEA-FOWL EGG LYSOZYMES
1JHL 1993-05-04T00:00:00+0000 2.4 THREE-DIMENSIONAL STRUCTURE OF A HETEROCLITIC ANTIGEN-ANTIBODY CROSS-REACTION COMPLEX
1BQL 1995-02-03T00:00:00+0000 2.6 STRUCTURE OF AN ANTI-HEL FAB FRAGMENT COMPLEXED WITH BOBWHITE QUAIL LYSOZYME
1DKJ 1996-01-10T00:00:00+0000 2.0 BOBWHITE QUAIL LYSOZYME
1DKK 1996-01-10T00:00:00+0000 1.9 BOBWHITE QUAIL LYSOZYME WITH NITRATE
135L 1993-06-10T00:00:00+0000 1.3 X-RAY STRUCTURE OF MONOCLINIC TURKEY EGG LYSOZYME AT 1.3 ANGSTROMS RESOLUTION

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
92.3 Lysozyme C P49663 LYSC_PHAVE
93.0 Lysozyme C P81711 LYSC_SYRSO
93.2 Lysozyme C P00702 LYSC_PHACO
94.6 Lysozyme C P24533 LYSC_SYRRE
93.0 Lysozyme C P24364 LYSC_LOPLE
92.2 Lysozyme C P00704 LYSC_NUMME
95.2 Lysozyme C P00703 LYSC_MELGA
95.3 Lysozyme C Q7LZT2 LYSC_TRATE
95.3 Lysozyme C P22910 LYSC_CHRAM
96.1 Lysozyme C P19849 LYSC_PAVCR
95.3 Lysozyme C P00701 LYSC_COTJA
96.1 Lysozyme C Q7LZI3 LYSC_TRASA
96.9 Lysozyme C Q7LZP9 LYSC_LOPIM
96.9 Lysozyme C Q7LZQ0 LYSC_CATWA
96.9 Lysozyme C P00699 LYSC_CALCC
96.9 Lysozyme C P00700 LYSC_COLVI
100.0 Lysozyme C P00698 LYSC_CHICK
96.0 Ribonuclease pancreatic Q29606 RNAS1_ORYLE
90.3 Ribonuclease pancreatic P00662 RNAS1_GIRCA
92.7 Ribonuclease pancreatic P00668 RNAS1_ANTAM
95.2 Ribonuclease pancreatic P00660 RNAS1_CONTA
93.5 Ribonuclease pancreatic P07848 RNAS1_EUDTH
96.0 Ribonuclease pancreatic P07847 RNAS1_AEPME
95.2 Ribonuclease pancreatic P00657 RNAS1_BUBBU
96.8 Ribonuclease pancreatic P67927 RNAS1_SHEEP
96.8 Ribonuclease pancreatic P67926 RNAS1_CAPHI
100.0 Ribonuclease pancreatic P61824 RNAS1_BISBI
100.0 Ribonuclease pancreatic P61823 RNAS1_BOVIN