Equilibrium and kinetic folding of hen egg-white lysozyme under acidic conditions.


Abstract

The equilibrium and kinetic folding of hen egg-white lysozyme was studied by means of circular dichroism spectra in the far- and near-ultraviolet (UV) regions at 25 degrees C under the acidic pH conditions. In equilibrium condition at pH 2.2, hen lysozyme shows a single cooperative transition in the GdnCl-induced unfolding experiment. However, in the GdnCl-induced unfolding process at lower pH 0.9, a distinct intermediate state with molten globule characteristics was observed. The time-dependent unfolding and refolding of the protein were induced by concentration jumps of the denaturant and measured by using stopped-flow circular dichroism at pH 2.2. Immediately after the dilution of denaturant, the kinetics of refolding shows evidence of a major unresolved far-UV CD change during the dead time (<10 ms) of the stopped-flow experiment (burst phase). The observed refolding and unfolding curves were both fitted well to a single-exponential function, and the rate constants obtained in the far- and near-UV regions coincided with each other. The dependence on denaturant concentration of amplitudes of burst phase and both rate constants was modeled quantitatively by a sequential three-state mechanism, U<-->I<-->N, in which the burst-phase intermediate (I) in rapid equilibrium with the unfolded state (U) precedes the rate-determining formation of the native state (N). The role of folding intermediate state of hen lysozyme was discussed. Study holds ProTherm entries: 17820, 17821, 17822, 17823 Extra Details: hen egg-white lysozyme; protein folding; stopped flow; folding intermediate

Submission Details

ID: 4dF7towi

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:50 p.m.

Version: 1

Publication Details
Sasahara K;Demura M;Nitta K,Proteins (2002) Equilibrium and kinetic folding of hen egg-white lysozyme under acidic conditions. PMID:12402357
Additional Information

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
100.0 Lysozyme C P00698 LYSC_CHICK
96.9 Lysozyme C P00700 LYSC_COLVI
96.9 Lysozyme C P00699 LYSC_CALCC
96.9 Lysozyme C Q7LZQ0 LYSC_CATWA
96.9 Lysozyme C Q7LZP9 LYSC_LOPIM
96.1 Lysozyme C Q7LZI3 LYSC_TRASA
95.3 Lysozyme C P00701 LYSC_COTJA
96.1 Lysozyme C P19849 LYSC_PAVCR
95.3 Lysozyme C P22910 LYSC_CHRAM
95.3 Lysozyme C Q7LZT2 LYSC_TRATE
95.2 Lysozyme C P00703 LYSC_MELGA
92.2 Lysozyme C P00704 LYSC_NUMME
93.0 Lysozyme C P24364 LYSC_LOPLE
94.6 Lysozyme C P24533 LYSC_SYRRE
93.2 Lysozyme C P00702 LYSC_PHACO
93.0 Lysozyme C P81711 LYSC_SYRSO
92.3 Lysozyme C P49663 LYSC_PHAVE