Domain structure and interactions of the type I and type II modules in the gelatin-binding region of fibronectin. All six modules are independently folded.


Abstract

The gelatin-binding region of fibronectin is isolated easily as a stable and functional 42 kDa fragment containing four type I "finger" modules and two type II "kringle-like" modules arranged in the order I6-II1-II2-I7-I8-I9. This fragment exhibits a single reversible melting transition near 64 degrees C in TBS buffer (0.02 M-Tris buffer containing 0.15 M-NaCl, pH 7.4). The transition is characterized by a calorimetric to van't Hoff enthalpy ratio of 1.6, suggesting a complex domain structure. A 30 kDa fragment with the same NH2 terminus (I6-II1-II2-I7) melts reversibly near 65 degrees C with delta Hcal/delta HvH = 1.3, also consistent with the presence of more than one domain. To elucidate further the domain structure, three non-overlapping subfragments were prepared and characterized with respect to their unfolding induced by heat and guanidinium chloride. The three subfragments, each containing two modules, are designated from amino or carboxyl-terminal location as 13 kDa (I6-II1) 16 kDa (II2-I7) and 21 kDa (I8-I9) according to their apparent Mr in SDS/polyacrylamide gel electrophoresis. All three subfragments exhibited reversible transitions in TBS buffer, behaving in the calorimeter as single co-operative units with delta Hcal/delta HvH close to unity. However, the specific enthalpies and changes in heat capacity associated with the melting of all fragments and subfragments in TBS buffer were low compared to those of most compact globular proteins, suggesting that not all modules are represented. When titrated with guanidinium chloride at 25 degrees C, all fragments exhibited monophasic reversible unfolding transitions detected by changes in fluorescence. Heating in the presence of 6 M-guanidinium chloride revealed three additional transitions not seen in the absence of denaturants. These transitions have been assigned to three of the four type I finger modules (I6, I7 and I9), one of which (I6) was isolated and shown to retain a compact structure as stable as that observed for this module within the parent fragments. Two other modules (II2 and I7) are destabilized when separated from their neighbors. Thus, despite their small size (50 to 60 amino acid residues), all six of the modules in the gelatin-binding region of fibronectin form independently folded domains, three of which (I6, I7 and I9) are unusually stable. Evidence is provided that four of the six modules interact with each other in the parent fragment. This interaction may explain previously noted disruptions in the otherwise uniform strand-like images seen in electron micrographs of fibronectin. Study holds ProTherm entries: 7124, 7125, 7126, 7127, 7128, 7129, 7130 Extra Details: gelatin-binding region; "kringle-like" modules; subfragments;,co-operative units

Submission Details

ID: RQo9nLvU3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:33 p.m.

Version: 1

Publication Details
Litvinovich SV;Strickland DK;Medved LV;Ingham KC,J. Mol. Biol. (1991) Domain structure and interactions of the type I and type II modules in the gelatin-binding region of fibronectin. All six modules are independently folded. PMID:1994038
Additional Information

Study Summary

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
1OWW 2003-10-07 Solution structure of the first type III module of human fibronectin determined by 1H, 15N NMR spectroscopy
2N1K 2015-11-11 Structure of the Third Type III Domain from Human Fibronectin
1E8B 2000-10-15 Solution structure of 6F11F22F2, a compact three-module fragment of the gelatin-binding domain of human fibronectin
2FN2 1998-09-16 SOLUTION NMR STRUCTURE OF THE GLYCOSYLATED SECOND TYPE TWO MODULE OF FIBRONECTIN, 20 STRUCTURES
1E88 2000-10-09 Solution structure of 6F11F22F2, a compact three-module fragment of the gelatin-binding domain of human fibronectin
1J8K 2002-02-06 NMR STRUCTURE OF THE FIBRONECTIN EDA DOMAIN, NMR, 20 STRUCTURES
2FNB 1998-12-23 NMR STRUCTURE OF THE FIBRONECTIN ED-B DOMAIN, NMR, 20 STRUCTURES
2CKU 2007-04-03 Solution structure of 2F13F1 from human fibronectin
1FBR 1995-10-15 FOURTH AND FIFTH FIBRONECTIN TYPE I MODULE PAIR
2EC3 2008-03-04 Solution structure of the 11th FN1 domain from human Fibronectin 1
5M0A 2017-11-29 Solution structure of isolated 15th Fibronectin III domain from human fibronectin
5J6Z 2017-04-12 Structure of anastellin bound to beta-strands A and B from the third type III domain of fibronectin
1TTF 1994-01-31 THE THREE-DIMENSIONAL STRUCTURE OF THE TENTH TYPE III MODULE OF FIBRONECTIN: AN INSIGHT INTO RGD-MEDIATED INTERACTIONS
1TTG 1994-01-31 THE THREE-DIMENSIONAL STRUCTURE OF THE TENTH TYPE III MODULE OF FIBRONECTIN: AN INSIGHT INTO RGD-MEDIATED INTERACTIONS
2HA1 2007-04-10 Complex of the first and second type III domains of human Fibronectin in solution
2H45 2007-04-10 Solution structure of the second type III domain of human Fibronectin: ensemble of 25 structures
1Q38 2003-11-04 Anastellin
1QO6 2000-01-11 Solution structure of a pair of modules from the gelatin-binding domain of fibronectin
1QGB 1999-12-08 SOLUTION STRUCTURE OF THE N-TERMINAL F1 MODULE PAIR FROM HUMAN FIBRONECTIN
2H41 2007-04-10 Solution structure of the second type III domain of human Fibronectin: minimized average structure
2MNU 2014-09-24 Backbone and side chain 1H, 13C, and 15N Chemical Shift Assignments for EDB and specific binding aptide
6HNF 2018-10-10 Structure in solution of human fibronectin type III-domain 14
1O9A 2003-05-08 Solution structure of the complex of 1F12F1 from fibronectin with B3 from FnBB from S. dysgalactiae
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
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
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
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
1FNA 1994-04-30 1.8 CRYSTAL STRUCTURE OF THE TENTH TYPE III CELL ADHESION MODULE OF HUMAN FIBRONECTIN
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
2CK2 2007-04-10 2.0 Structure of core-swapped mutant of fibronectin
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
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
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