Urea-induced sequential unfolding of fibronectin: a fluorescence spectroscopy and circular dichroism study.


Abstract

Fibronectin (FN) is an extracellular matrix (ECM) protein found soluble in corporal fluids or as an insoluble fibrillar component incorporated in the ECM. This phenomenon implicates structural changes that expose FN binding sites and activate the protein to promote intermolecular interactions with other FN. We have investigated, using fluorescence and circular dichroism spectroscopy, the unfolding process of human fibronectin induced by urea in different ionic strength conditions. At any ionic strength, the equilibrium unfolding data are well described by a four-state equilibrium model N <= => I(1) <= =>I(2) <= => U. Fitting this model to experimental values, we have determined the free energy change for the different steps. We found that the N <= => I(1) transition corresponds to a free energy of 10.5 +/- 0.4 kcal/mol. Comparable values of free energy change are generally associated with a partial unfolding of the type III domain. For the I(1) <= => I(2) transition, the free energy change is 7.6 +/- 0.4 kcal/mol at low ionic strength but is twice as low at high ionic strength. This result is consistent with observations indicating that the complete unfolding of the type III domain from partially unfolded forms necessitates about 5 kcal/mol. The third step, I(2) <= => U, which leads to the complete unfolding of fibronectin, corresponds to a free energy change of 14.4 +/- 0.9 kcal/mol at low ionic strength whereas this energy is again twice as low under high ionic strength conditions. This hierarchical unfolding of fibronectin, as well as the stability of the different intermediates controlled by ionic strength demonstrated here, could be important for the understanding of activation of the matrix assembly. Study holds ProTherm entries: 16863, 16864, 16865, 16866, 16867, 16868, 16869, 16870, 16871, 16872, 16873, 16874, 16875, 16876, 16877 Extra Details: Transition 1. Experiment was done in pure water, 18 M ohm. structural changes; binding sites; intermolecular interactions; partial unfolding; ionic strength

Submission Details

ID: P9sMUN5C4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:48 p.m.

Version: 1

Publication Details
Patel S;Chaffotte AF;Goubard F;Pauthe E,Biochemistry (2004) Urea-induced sequential unfolding of fibronectin: a fluorescence spectroscopy and circular dichroism study. PMID:14769050
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
2H45 2007-04-10 Solution structure of the second type III domain of human Fibronectin: ensemble of 25 structures
6HNF 2018-10-10 Structure in solution of human fibronectin type III-domain 14
2FN2 1998-09-16 SOLUTION NMR STRUCTURE OF THE GLYCOSYLATED SECOND TYPE TWO MODULE OF FIBRONECTIN, 20 STRUCTURES
5J6Z 2017-04-12 Structure of anastellin bound to beta-strands A and B from the third type III domain of fibronectin
2EC3 2008-03-04 Solution structure of the 11th FN1 domain from human Fibronectin 1
1E88 2000-10-09 Solution structure of 6F11F22F2, a compact three-module fragment of the gelatin-binding domain of human fibronectin
2H41 2007-04-10 Solution structure of the second type III domain of human Fibronectin: minimized average structure
1TTG 1994-01-31 THE THREE-DIMENSIONAL STRUCTURE OF THE TENTH TYPE III MODULE OF FIBRONECTIN: AN INSIGHT INTO RGD-MEDIATED INTERACTIONS
1OWW 2003-10-07 Solution structure of the first type III module of human fibronectin determined by 1H, 15N NMR spectroscopy
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
1FBR 1995-10-15 FOURTH AND FIFTH FIBRONECTIN TYPE I MODULE PAIR
2HA1 2007-04-10 Complex of the first and second type III domains of human Fibronectin in solution
2N1K 2015-11-11 Structure of the Third Type III Domain from Human Fibronectin
2MNU 2014-09-24 Backbone and side chain 1H, 13C, and 15N Chemical Shift Assignments for EDB and specific binding aptide
1E8B 2000-10-15 Solution structure of 6F11F22F2, a compact three-module fragment of the gelatin-binding domain of human fibronectin
1O9A 2003-05-08 Solution structure of the complex of 1F12F1 from fibronectin with B3 from FnBB from S. dysgalactiae
1J8K 2002-02-06 NMR STRUCTURE OF THE FIBRONECTIN EDA DOMAIN, NMR, 20 STRUCTURES
2CKU 2007-04-03 Solution structure of 2F13F1 from human fibronectin
5M0A 2017-11-29 Solution structure of isolated 15th Fibronectin III domain from human fibronectin
2FNB 1998-12-23 NMR STRUCTURE OF THE FIBRONECTIN ED-B DOMAIN, NMR, 20 STRUCTURES
1QGB 1999-12-08 SOLUTION STRUCTURE OF THE N-TERMINAL F1 MODULE PAIR FROM HUMAN FIBRONECTIN
1QO6 2000-01-11 Solution structure of a pair of modules from the gelatin-binding domain of 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
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
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
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
5DFT 2016-09-14 2.5 Structure of the Eleventh Type III Domain from Human Fibronectin
3M7P 2010-06-23 2.5 Fibronectin fragment
5J7C 2016-08-17 2.54 A picomolar affinity FN3 domain in complex with hen egg-white lysozyme
3GXE 2010-04-07 2.6 Complex of a Low Affinity Collagen Site with the Fibronectin 8-9FnI Domain Pair
4GH7 2012-12-26 2.6 Crystal structure of Anticalin N7A in complex with oncofetal fibronectin fragment Fn7B8
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
3MQL 2010-08-25 3.0 Crystal structure of the fibronectin 6FnI1-2FnII7FnI fragment
5N47 2018-02-14 3.0 Structure of Anticalin N7E in complex with the three-domain fragment Fn7B8 of human oncofetal fibronectin
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