Role of the N-terminal helix I for dimerization and stability of the calcium-binding protein S100B.


Abstract

Human S100B(beta beta) is a small intracellular EF-hand calcium-binding protein that consists of two noncovalently associated 91-residue beta monomers. The three-dimensional structures of S100B reveal the dimer interface consists of four alpha-helices (I, I' and IV, IV') packed in an X-type bundle. In this study, guanidine hydrochloride denaturation and dynamic light scattering were used to assess the impact of single (L3A, L3S, M7A, I11A, F14A) and double (L3A/I11A and L3A/F14A) substitution mutations in helix I on the stability and dimerization propensity of S100B. The free energy of unfolding (Delta G(u)) of wild-type apo-S100B was determined to be 72.4 +/- 4.0 kJ mol(-1), consistent with it being the most stable calcium-binding protein to date. The order of stability of the mutants in their apo form is S100B > L3A > L3S > I11A > M7A approximately L3A/I11A > F14A > L3A/F14A. Further, there is a strong correlation between the stability and the cooperativity of unfolding. Each mutation proved to be more stable in its calcium form compared to its apo form. The calcium-bound L3S substitution proved to be significantly more stable than calcium-saturated S100B, whereas the L3A, I11A, and L3A/I11A mutants are only slightly more stable than the wild-type protein. The F14A and L3A/F14A mutants are significantly reduced in stability, even in the presence of calcium. Study holds ProTherm entries: 13045, 13046, 13047, 13048, 13049, 13050, 13051, 13052, 13053, 13054, 13055, 13056, 13057, 13058, 13059, 13060 Extra Details: apo form; 1mM DTT and 1mM EDTA were added in the experiment. EF-hand; free energy; cooperativity; calcium

Submission Details

ID: RgpQ7zPQ3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:44 p.m.

Version: 1

Publication Details
Ferguson PL;Shaw GS,Biochemistry (2002) Role of the N-terminal helix I for dimerization and stability of the calcium-binding protein S100B. PMID:11888280
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
1DT7 2000-07-26 SOLUTION STRUCTURE OF THE C-TERMINAL NEGATIVE REGULATORY DOMAIN OF P53 IN A COMPLEX WITH CA2+-BOUND S100B(BB)
1MWN 2002-12-18 Solution NMR structure of S100B bound to the high-affinity target peptide TRTK-12
2PRU 2008-04-15 NMR Structure of Human apoS100B at 10C
1MQ1 2002-12-25 Ca2+-S100B-TRTK-12 complex
1QLK 1998-11-11 SOLUTION STRUCTURE OF CA(2+)-LOADED RAT S100B (BETABETA) NMR, 20 STRUCTURES
1CFP 1997-03-12 S100B (S100BETA) NMR DATA WAS COLLECTED FROM A SAMPLE OF CALCIUM FREE PROTEIN AT PH 6.3 AND A TEMPERATURE OF 311 K AND 1.7-6.9 MM CONCENTRATION, 25 STRUCTURES
1PSB 2003-12-16 Solution structure of calcium loaded S100B complexed to a peptide from N-Terminal regulatory domain of NDR kinase.
2M49 2013-12-18 Structural Insights into Human S100B and Basic Fibroblast Growth Factor (FGF2) Interaction
2K7O 2008-11-18 Ca2+-S100B, refined with RDCs
1UWO 1998-06-10 CALCIUM FORM OF HUMAN S100B, NMR, 20 STRUCTURES
1B4C 1998-12-30 SOLUTION STRUCTURE OF RAT APO-S100B USING DIPOLAR COUPLINGS
1XYD 2005-06-07 NMR Solution Structure of Rat Zinc-Calcium-S100B, 20 Structures
1SYM 1996-12-07 3-D SOLUTION STRUCTURE OF REDUCED APO-S100B FROM RAT, NMR, 20 STRUCTURES
4PE0 2014-11-05 1.08 Crystal Structure of Calcium-loaded S100B bound to SBi4434
3RM1 2012-04-25 1.24 1.24 Angstrom X-ray structure of bovine TRTK12-Ca(2+)-S100B D63N
5ER5 2016-06-08 1.26 Crystal Structure of Calcium-loaded S100B bound to SC1990
5D7F 2016-08-17 1.3 X-ray structure of Ca(2+)-S100B with human RAGE-derived W72 peptide
3CZT 2009-04-14 1.4 Crystal Structure of S100B in the Calcium and Zinc Loaded State at pH 9
3IQO 2010-02-02 1.5 1.5 angstrom X-ray structure of bovine Ca(2+)-S100B
3D0Y 2009-04-14 1.5 Crystal Structure of S100B in the Calcium and Zinc Loaded State at pH 6.5
5DKN 2016-01-20 1.53 Crystal Structure of Calcium-loaded S100B bound to SBi4225
4PE1 2014-10-15 1.58 Crystal Structure of Calcium-loaded S100B bound to SC124
5DKQ 2016-01-20 1.59 Crystal Structure of Calcium-loaded S100B bound to SBi4214
4FQO 2012-10-17 1.65 Crystal Structure of Calcium-Loaded S100B Bound to SBi4211
4PE7 2014-10-15 1.65 Crystal Structure of Calcium-loaded S100B bound to SC1982
3D10 2009-04-14 1.65 Crystal Structure of S100B in the Calcium and Zinc Loaded State at pH 10.0
4PDZ 2014-10-15 1.73 Crystal Structure of Calcium-loaded S100B bound to SBi4172
5DKR 2016-01-20 1.74 Crystal Structure of Calcium-loaded S100B bound to SBi29
3LK1 2010-12-29 1.79 X-ray structure of bovine SC0322,Ca(2+)-S100B
5ER4 2016-06-08 1.81 Crystal Structure of Calcium-loaded S100B bound to SC0025
3LLE 2010-12-29 1.85 X-ray structure of bovine SC0322,Ca(2+)-S100B
3CR5 2008-06-24 1.85 X-ray structure of bovine Pnt-Zn(2+),Ca(2+)-S100B
3CR2 2008-06-24 1.88 X-ray structure of bovine Zn(2+),Ca(2+)-S100B
2H61 2007-06-05 1.9 X-ray structure of human Ca2+-loaded S100B
3GK4 2009-06-09 1.9 X-ray structure of bovine SBi523,Ca(2+)-S100B
3GK2 2009-06-09 1.98 X-ray structure of bovine SBi279,Ca(2+)-S100B
3HCM 2010-02-02 2.0 Crystal structure of human S100B in complex with S45
1MHO 1998-11-18 2.0 THE 2.0 A STRUCTURE OF HOLO S100B FROM BOVINE BRAIN
3RLZ 2012-04-25 2.01 2.0 Angstrom X-ray structure of bovine Ca(2+)-S100B D63N
3IQQ 2010-02-02 2.01 X-ray structure of bovine TRTK12-Ca(2+)-S100B
3LK0 2010-12-29 2.04 X-ray structure of bovine SC0067,Ca(2+)-S100B
3GK1 2009-06-09 2.1 X-ray structure of bovine SBi132,Ca(2+)-S100B
5CSI 2015-11-11 2.13 S100B-RSK1 crystal structure A'
3CR4 2008-08-05 2.15 X-ray structure of bovine Pnt,Ca(2+)-S100B
4PE4 2014-10-15 2.18 Crystal Structure of Calcium-loaded S100B bound to SC1475
5CSF 2015-11-11 2.4 S100B-RSK1 crystal structure A
4XYN 2015-05-13 2.55 X-ray structure of Ca(2+)-S100B with human RAGE-derived W61 peptide
5CSJ 2015-11-11 2.7 S100B-RSK1 crystal structure B
5CSN 2015-11-11 2.95 S100B-RSK1 crystal structure C

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
96.7 Protein S100-B P02638 S100B_BOVIN
97.8 Protein S100-B P04631 S100B_RAT
98.9 Protein S100-B P50114 S100B_MOUSE
100.0 Protein S100-B Q6YNR6 S100B_RABIT
100.0 Protein S100-B P04271 S100B_HUMAN