Folding and stability of a fibronectin type III domain of human tenascin.


Abstract

The folding of an isolated fibronectin type III domain of human tenascin, a large extra-cellular matrix protein, has been characterised. The isolated module, which has no disulphide bonds, can be reversibly unfolded by chemical denaturant and temperature. Equilibrium unfolding, measured using a number of different probes, fits to a two-state transition, with consistent measures of DeltaGH2OD-N. Folding and refolding rate constants have been determined over a range of denaturant concentrations. The refolding kinetics are bi-phasic, and in the transition region the slow phase dominates refolding kinetics. Outside the transition region the folding of the fast-folding species fits to a two-state model. There is no evidence for significant accumulation of partially folded intermediates. Study holds ProTherm entries: 8887, 8888, 8889, 8890, 8891, 8892, 8893, 8894, 8895, 8896 Extra Details: Fluorescence measurements at 350nm tenascin; fibronectin type III; immunoglobulin; protein folding; stability

Submission Details

ID: xfkYQ7rA4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:37 p.m.

Version: 1

Publication Details
Clarke J;Hamill SJ;Johnson CM,J. Mol. Biol. (1997) Folding and stability of a fibronectin type III domain of human tenascin. PMID:9245604
Additional Information

Study Summary

Number of data points 22
Proteins Fibronectin ; Fibronectin
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: Tm pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: Cm pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: m pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: dG_H2O pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: Cm prot_conc:1 micro M, buffers:Sodium phosphate: 50 mM, pH:7.0 ; Experimental Assay: m prot_conc:1 micro M, buffers:Sodium phosphate: 50 mM, pH:7.0 ; Experimental Assay: dG_H2O prot_conc:1 micro M, buffers:Sodium phosphate: 50 mM, pH:7.0 ; Experimental Assay: Cm pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:25 micro M ; Experimental Assay: m pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:25 micro M ; Experimental Assay: dG_H2O pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:25 micro M ; Experimental Assay: Tm pH:5.0, prot_conc:4 micro MM, buffers:Sodium acetate: 50 mM ; Experimental Assay: Tm prot_conc:1 micro M, pH:5.0, buffers:Sodium acetate: 50 mM ; Experimental Assay: Cm prot_conc:0.12 mg/ml, pH:5.0, buffers:Sodium acetate: 50 mM ; Experimental Assay: m prot_conc:0.12 mg/ml, pH:5.0, buffers:Sodium acetate: 50 mM ; Experimental Assay: dG_H2O prot_conc:0.12 mg/ml, pH:5.0, buffers:Sodium acetate: 50 mM
Libraries Mutations for sequence VSDVPRDLEVVAATPTSLLISWDAPAVTVRYYRITYGETGGNSPVQEFTVPGSKSTATISGLKPGVDYTITVYAVTGRGDSPASSKPISINYRT

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
1J8K 2002-02-06 NMR STRUCTURE OF THE FIBRONECTIN EDA DOMAIN, NMR, 20 STRUCTURES
6HNF 2018-10-10 Structure in solution of human fibronectin type III-domain 14
2FNB 1998-12-23 NMR STRUCTURE OF THE FIBRONECTIN ED-B DOMAIN, NMR, 20 STRUCTURES
2N1K 2015-11-11 Structure of the Third Type III Domain from Human Fibronectin
2HA1 2007-04-10 Complex of the first and second type III domains of human Fibronectin in solution
1E8B 2000-10-15 Solution structure of 6F11F22F2, a compact three-module fragment of the gelatin-binding domain of human fibronectin
1TTG 1994-01-31 THE THREE-DIMENSIONAL STRUCTURE OF THE TENTH TYPE III MODULE OF FIBRONECTIN: AN INSIGHT INTO RGD-MEDIATED INTERACTIONS
2H45 2007-04-10 Solution structure of the second type III domain of human Fibronectin: ensemble of 25 structures
1OWW 2003-10-07 Solution structure of the first type III module of human fibronectin determined by 1H, 15N NMR spectroscopy
2MNU 2014-09-24 Backbone and side chain 1H, 13C, and 15N Chemical Shift Assignments for EDB and specific binding aptide
5M0A 2017-11-29 Solution structure of isolated 15th Fibronectin III domain from human fibronectin
1Q38 2003-11-04 Anastellin
1TTF 1994-01-31 THE THREE-DIMENSIONAL STRUCTURE OF THE TENTH TYPE III MODULE OF FIBRONECTIN: AN INSIGHT INTO RGD-MEDIATED INTERACTIONS
1O9A 2003-05-08 Solution structure of the complex of 1F12F1 from fibronectin with B3 from FnBB from S. dysgalactiae
5J6Z 2017-04-12 Structure of anastellin bound to beta-strands A and B from the third type III domain of fibronectin
2H41 2007-04-10 Solution structure of the second type III domain of human Fibronectin: minimized average structure
2FN2 1998-09-16 SOLUTION NMR STRUCTURE OF THE GLYCOSYLATED SECOND TYPE TWO MODULE OF FIBRONECTIN, 20 STRUCTURES
2CKU 2007-04-03 Solution structure of 2F13F1 from human fibronectin
2EC3 2008-03-04 Solution structure of the 11th FN1 domain from human Fibronectin 1
1QO6 2000-01-11 Solution structure of a pair of modules from the gelatin-binding domain of fibronectin
1FBR 1995-10-15 FOURTH AND FIFTH FIBRONECTIN TYPE I MODULE PAIR
1E88 2000-10-09 Solution structure of 6F11F22F2, a compact three-module fragment of the gelatin-binding domain of human fibronectin
1QGB 1999-12-08 SOLUTION STRUCTURE OF THE N-TERMINAL F1 MODULE PAIR FROM HUMAN FIBRONECTIN
2CG7 2007-02-27 1.2 SECOND AND THIRD FIBRONECTIN TYPE I MODULE PAIR (CRYSTAL FORM II).
4LXO 2014-08-06 1.42 Crystal structure of 9,10Fn3-elegantin chimera
5DC4 2016-03-02 1.48 CRYSTAL STRUCTURE OF MONOBODY AS25/ABL1 SH2 DOMAIN COMPLEX, CRYSTAL A
2CG6 2007-02-27 1.55 Second and third fibronectin type I module pair (crystal form I).
5DC9 2016-03-02 1.56 CRYSTAL STRUCTURE OF MONOBODY AS25/ABL1 SH2 DOMAIN COMPLEX, CRYSTAL B
5N48 2018-02-14 1.6 Structure of Anticalin N9B in complex with extra-domain B of human oncofetal fibronectin
3CAL 2008-08-05 1.7 Crystal structure of the second and third fibronectin F1 modules in complex with a fragment of staphylococcus aureus fnbpa-5
3ZRZ 2011-09-28 1.7 Crystal structure of the second and third fibronectin F1 modules in complex with a fragment of Streptococcus pyogenes SfbI-5
1FNA 1994-04-30 1.8 CRYSTAL STRUCTURE OF THE TENTH TYPE III CELL ADHESION MODULE OF HUMAN FIBRONECTIN
2RKY 2008-08-05 1.8 Crystal structure of the fourth and fifth fibronectin F1 modules in complex with a fragment of staphylococcus aureus fnbpa-1
4PZ5 2014-07-02 1.96 Crystal structure of the second and third fibronectin F1 modules in complex with a fragment of BBK32 from Borrelia burgdorferi
2RL0 2008-08-05 2.0 Crystal structure of the fourth and fifth fibronectin F1 modules in complex with a fragment of staphylococcus aureus fnbpa-5
2RKZ 2008-08-05 2.0 Crystal structure of the second and third fibronectin f1 modules in complex with a fragment of staphylococcus aureus fnbpa-1
1FNF 1996-01-29 2.0 FRAGMENT OF HUMAN FIBRONECTIN ENCOMPASSING TYPE-III REPEATS 7 THROUGH 10
2CK2 2007-04-10 2.0 Structure of core-swapped mutant of fibronectin
2GEE 2006-10-17 2.01 Crystal Structure of Human Type III Fibronectin Extradomain B and Domain 8
3EJH 2009-02-03 2.1 Crystal Structure of the Fibronectin 8-9FnI Domain Pair in Complex with a Type-I Collagen Peptide
5DC0 2016-03-02 2.23 CRYSTAL STRUCTURE OF MONOBODY GG3/ABL1 SH2 DOMAIN COMPLEX
4JEG 2013-08-28 2.3 Crystal Structure of Monobody CS1/SHP2 C-SH2 Domain Complex
4JE4 2013-08-28 2.31 Crystal Structure of Monobody NSa1/SHP2 N-SH2 Domain Complex
3T1W 2012-03-21 2.4 Structure of the four-domain fragment Fn7B89 of oncofetal fibronectin
3R8Q 2012-04-11 2.4 Structure of Fibronectin domain 12-14
3M7P 2010-06-23 2.5 Fibronectin fragment
5DFT 2016-09-14 2.5 Structure of the Eleventh Type III Domain from Human Fibronectin
5J7C 2016-08-17 2.54 A picomolar affinity FN3 domain in complex with hen egg-white lysozyme
4GH7 2012-12-26 2.6 Crystal structure of Anticalin N7A in complex with oncofetal fibronectin fragment Fn7B8
3GXE 2010-04-07 2.6 Complex of a Low Affinity Collagen Site with the Fibronectin 8-9FnI Domain Pair
1FNH 1999-03-16 2.8 CRYSTAL STRUCTURE OF HEPARIN AND INTEGRIN BINDING SEGMENT OF HUMAN FIBRONECTIN
2OCF 2007-11-06 2.95 Human estrogen receptor alpha ligand-binding domain in complex with estradiol and the E2#23 FN3 monobody
5N47 2018-02-14 3.0 Structure of Anticalin N7E in complex with the three-domain fragment Fn7B8 of human oncofetal fibronectin
3MQL 2010-08-25 3.0 Crystal structure of the fibronectin 6FnI1-2FnII7FnI fragment
4MMZ 2014-03-26 3.1 Integrin AlphaVBeta3 ectodomain bound to an antagonistic tenth domain of Fibronectin
4MMY 2014-03-26 3.18 Integrin AlphaVBeta3 ectodomain bound to the tenth domain of Fibronectin with the IAKGDWND motif
4MMX 2014-03-26 3.32 Integrin AlphaVBeta3 ectodomain bound to the tenth domain of Fibronectin

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
96.8 Fibronectin P07589 FINC_BOVIN
100.0 Fibronectin P02751 FINC_HUMAN
90.9 Fibronectin Q91400 FINC_NOTVI
92.1 Fibronectin Q28377 FINC_HORSE
94.0 Fibronectin Q28275 FINC_CANLF