Gamma S-crystallin of bovine and human eye lens: solution structure, stability and folding of the intact two-domain protein and its separate domains.


Abstract

Human and bovine gammaS-crystallin (HgammaS and BgammaS) and their isolated N- and C-terminal domains were cloned and expressed as recombinant proteins in E. coli. HgammaS and BgammaS are found to be authentic according to their spectral and hydrodynamic properties. Both full-length proteins and isolated domains are monomeric and exhibit high thermal and pH stabilities. The thermodynamic characterization made use of chemically and thermally-induced equilibrium unfolding transitions at varying pH. In spite of its exemplary two-domain structure, gammaS-crystallin does not show bimodal unfolding characteristics. In the case of BgammaS, at pH 7.0, the C-terminal domain is less stable than the N-terminal one, whereas for HgammaS the opposite holds true. Differential scanning calorimetry confirms the results of chemically-induced equilibrium unfolding transitions. Over the whole pH range between 2.0 and 11.5, HgammaS-crystallin and its isolated domains (HgammaS-N and HgammaS-C) follow the two-state model. The two-state unfolding of the intact two-domain protein points to the close similarity of the stabilities of the constituent domains. Obviously, interactions between the domains do not contribute significantly to the overall stability of gammaS-crystallin. In contrast, the structurally closely related gammaB-crystallin owes much of its extreme stability to domain interactions. Study holds ProTherm entries: 12007, 12008, 12009 Extra Details: additive : DTT(5 mM), crystallins; differential scanning calorimetry; protein folding;,protein stability; domain interaction

Submission Details

ID: ey8Too7d3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:43 p.m.

Version: 1

Publication Details
Wenk M;Herbst R;Hoeger D;Kretschmar M;Lubsen NH;Jaenicke R,Biophys. Chem. (2000) Gamma S-crystallin of bovine and human eye lens: solution structure, stability and folding of the intact two-domain protein and its separate domains. PMID:11026675
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
1ZWO 2005-07-05 NMR structure of murine gamma-S crystallin
6IF9 2019-04-10 Solution-state NMR structure of G57W human gammaS crystallin
2M3T 2013-11-13 Solution-state NMR structure of wild-type human gamma(S)-crystallin
2M3U 2013-11-13 Solution-state NMR structure of cataract-related human gamma(S)-crystallin point variant G18V
2A5M 2005-07-19 NMR structure of murine gamma-S crystallin from joint refinement with SAXS data
1ZWM 2005-07-05 NMR structure of murine gamma-S crystallin
6FD8 2018-12-26 2.1 Gamma-s crystallin dimer
1HA4 2001-11-20 2.4 GammaS crystallin C terminal domain from Homo Sapiens
1A7H 1998-05-27 2.56 GAMMA S CRYSTALLIN C-TERMINAL DOMAIN
6MYH 2018-11-14 2.9 Mouse Gamma S Crystallin L16 Octamer
6MYG 2018-11-14 2.92 Mouse Gamma S Crystallin L16 Octamer

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
92.0 Beta-crystallin S P0C5E9 CRYGS_RAT
90.8 Beta-crystallin S O35486 CRYGS_MOUSE
95.4 Beta-crystallin S A2IBY7 CRYGS_CANLF
93.1 Beta-crystallin S P06504 CRYGS_BOVIN
94.3 Beta-crystallin S A4L9I8 CRYGS_RABIT
100.0 Beta-crystallin S P22914 CRYGS_HUMAN