Effect of dextran on protein stability and conformation attributed to macromolecular crowding.


Abstract

Thermally induced transition curves of hen egg-white lysozyme were measured in the presence of several concentrations of dextran at pH 2.0 by near-UV and far-UV CD. The transition curves were fitted to a two-state model by a non-linear, least-squares method to obtain the transition temperature (T(m)), enthalpy change (deltaH(u)(T(m))), and free energy change (deltaG(u)(T)) of the unfolding transition. An increase in T(m) and almost constant deltaH(u)(T(m)) values were observed in the presence of added dextran at concentrations exceeding ca 100 g l(-1). In addition, dextran-induced conformational changes of fully unfolded protein were investigated by CD spectroscopy. Addition of high concentrations of dextran to solutions of acid-unfolded cytochrome c at pH 2.0 results in a shift of the CD spectrum from that characteristic of the fully unfolded polypeptide to that characteristic of the more compact, salt-induced molten globule state, a result suggesting that the molten globule-like state is stabilized relative to the fully unfolded form in crowded environments. Both observations are in qualitative accord with predictions of a previously proposed model for the effect of intermolecular excluded volume (macromolecular crowding) on protein stability and conformation. Study holds ProTherm entries: 16230, 16231, 16232, 16233, 16234, 16235, 16236, 16237, 16238, 16239, 16240, 16241, 16242, 16243, 16244, 16245 Extra Details: (i) at 289 nm protein stability; molecular crowding; molten globule; excluded volume; dextran

Submission Details

ID: PCSoUTst

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:47 p.m.

Version: 1

Publication Details
Sasahara K;McPhie P;Minton AP,J. Mol. Biol. (2003) Effect of dextran on protein stability and conformation attributed to macromolecular crowding. PMID:12589765
Additional Information

Study Summary

Number of data points 48
Proteins Lysozyme C ; Lysozyme C
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: dCp prot_conc:0.78 g/l, details:Additives dextran (300 g/L), ; Experimental Assay: Tm prot_conc:0.78 g/l, details:Additives dextran (300 g/L), ; Experimental Assay: dHvH prot_conc:0.78 g/l, details:Additives dextran (300 g/L), ; Experimental Assay: dCp details:Additives dextran (300 g/L),, prot_conc:0.41 g/l ; Experimental Assay: Tm details:Additives dextran (300 g/L),, prot_conc:0.41 g/l ; Experimental Assay: dHvH details:Additives dextran (300 g/L),, prot_conc:0.41 g/l ; Experimental Assay: dCp prot_conc:0.21 g/l, details:Additives dextran (300 g/L), ; Experimental Assay: Tm prot_conc:0.21 g/l, details:Additives dextran (300 g/L), ; Experimental Assay: dHvH prot_conc:0.21 g/l, details:Additives dextran (300 g/L), ; Experimental Assay: dCp details:Additives dextran (250 g/L),, prot_conc:0.38 g/l ; Experimental Assay: Tm details:Additives dextran (250 g/L),, prot_conc:0.38 g/l ; Experimental Assay: dHvH details:Additives dextran (250 g/L),, prot_conc:0.38 g/l ; Experimental Assay: dCp details:Additives dextran (190 g/L),, prot_conc:0.39 g/l ; Experimental Assay: Tm details:Additives dextran (190 g/L),, prot_conc:0.39 g/l ; Experimental Assay: dHvH details:Additives dextran (190 g/L),, prot_conc:0.39 g/l ; Experimental Assay: dCp details:Additives dextran (150 g/L),, prot_conc:0.38 g/l ; Experimental Assay: Tm details:Additives dextran (150 g/L),, prot_conc:0.38 g/l ; Experimental Assay: dHvH details:Additives dextran (150 g/L),, prot_conc:0.38 g/l ; Experimental Assay: dCp details:Additives dextran (100 g/L),, prot_conc:0.39 g/l ; Experimental Assay: Tm details:Additives dextran (100 g/L),, prot_conc:0.39 g/l ; Experimental Assay: dHvH details:Additives dextran (100 g/L),, prot_conc:0.39 g/l ; Experimental Assay: dCp details:Additives , prot_conc:0.39 g/l ; Experimental Assay: Tm details:Additives , prot_conc:0.39 g/l ; Experimental Assay: dHvH details:Additives , prot_conc:0.39 g/l
Libraries 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)
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
1DZB 2000-02-23T00:00:00+0000 2.0 Crystal structure of phage library-derived single-chain Fv fragment 1F9 in complex with turkey egg-white lysozyme

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