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Implications for collagen binding from the crystallographic structure of fibronectin 6FnI1-2FnII7FnI.

Erat MC, Schwarz-Linek U, Pickford AR, Farndale RW, Campbell ID, Vakonakis I - J. Biol. Chem. (2010)

Bottom Line: Analysis of NMR titrations with single-stranded collagen peptides reveals a dominant collagen interaction surface on domains (2)FnII and (7)FnI; a similar surface appears involved in interactions with triple-helical peptides.Models of the complete GBD, based on the new structure and the (8-9)FnI·collagen complex show a continuous putative collagen binding surface.We explore the implications of this model using long collagen peptides and discuss our findings in the context of FN interactions with collagen fibrils.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.

ABSTRACT
Collagen and fibronectin (FN) are two abundant and essential components of the vertebrate extracellular matrix; they interact directly with cellular receptors and affect cell adhesion and migration. Past studies identified a FN fragment comprising six modules, (6)FnI(1-2)FnII(7-9)FnI, and termed the gelatin binding domain (GBD) as responsible for collagen interaction. Recently, we showed that the GBD binds tightly to a specific site within type I collagen and determined the structure of domains (8-9)FnI in complex with a peptide from that site. Here, we present the crystallographic structure of domains (6)FnI(1-2)FnII(7)FnI, which form a compact, globular unit through interdomain interactions. Analysis of NMR titrations with single-stranded collagen peptides reveals a dominant collagen interaction surface on domains (2)FnII and (7)FnI; a similar surface appears involved in interactions with triple-helical peptides. Models of the complete GBD, based on the new structure and the (8-9)FnI·collagen complex show a continuous putative collagen binding surface. We explore the implications of this model using long collagen peptides and discuss our findings in the context of FN interactions with collagen fibrils.

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Prominent domain-domain contacts involving 6FnI-2FnII (A), 1FnII-2FnII (B), and 1–2FnII-7FnI (C). Individual domains are colored as in Fig. 1, and specific residues are shown as sticks colored similar to their respective domains. Compared with the solution structure of 6FnI1–2FnII (12) a new 1FnII-2FnII interface (B) is formed, and the 2FnII-7FnI linker is stabilized. 7FnI interacts with both 1FnII and 2FnII and is further anchored to this linker.
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Figure 2: Prominent domain-domain contacts involving 6FnI-2FnII (A), 1FnII-2FnII (B), and 1–2FnII-7FnI (C). Individual domains are colored as in Fig. 1, and specific residues are shown as sticks colored similar to their respective domains. Compared with the solution structure of 6FnI1–2FnII (12) a new 1FnII-2FnII interface (B) is formed, and the 2FnII-7FnI linker is stabilized. 7FnI interacts with both 1FnII and 2FnII and is further anchored to this linker.

Mentions: The compact 6FnI1–2FnII7FnI conformation is maintained through interdomain interactions that show a remarkable degree of conservation (supplemental Fig. 1). 6FnI interacts with 2FnII in a manner essentially identical to that observed in the solution 6FnI1–2FnII structure (12), burying ∼395 Å2 of solvent-accessible surface area (Fig. 2A). The Cα root mean square deviation for 6FnI·2FnII between the solution structure and our crystallographic model is only 1.7 Å, whereas the individual domains differ by 1.0 and 1.1 Å for 6FnI and 2FnII, respectively. Residues 314–323 of 6FnI, and 414–421 and 448–449 of 2FnII are primarily involved in forming the interface, with significant contributions from Met320, Ser415, Ala418, Leu419, Thr448, and Thr449 (Fig. 2A).


Implications for collagen binding from the crystallographic structure of fibronectin 6FnI1-2FnII7FnI.

Erat MC, Schwarz-Linek U, Pickford AR, Farndale RW, Campbell ID, Vakonakis I - J. Biol. Chem. (2010)

Prominent domain-domain contacts involving 6FnI-2FnII (A), 1FnII-2FnII (B), and 1–2FnII-7FnI (C). Individual domains are colored as in Fig. 1, and specific residues are shown as sticks colored similar to their respective domains. Compared with the solution structure of 6FnI1–2FnII (12) a new 1FnII-2FnII interface (B) is formed, and the 2FnII-7FnI linker is stabilized. 7FnI interacts with both 1FnII and 2FnII and is further anchored to this linker.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2962475&req=5

Figure 2: Prominent domain-domain contacts involving 6FnI-2FnII (A), 1FnII-2FnII (B), and 1–2FnII-7FnI (C). Individual domains are colored as in Fig. 1, and specific residues are shown as sticks colored similar to their respective domains. Compared with the solution structure of 6FnI1–2FnII (12) a new 1FnII-2FnII interface (B) is formed, and the 2FnII-7FnI linker is stabilized. 7FnI interacts with both 1FnII and 2FnII and is further anchored to this linker.
Mentions: The compact 6FnI1–2FnII7FnI conformation is maintained through interdomain interactions that show a remarkable degree of conservation (supplemental Fig. 1). 6FnI interacts with 2FnII in a manner essentially identical to that observed in the solution 6FnI1–2FnII structure (12), burying ∼395 Å2 of solvent-accessible surface area (Fig. 2A). The Cα root mean square deviation for 6FnI·2FnII between the solution structure and our crystallographic model is only 1.7 Å, whereas the individual domains differ by 1.0 and 1.1 Å for 6FnI and 2FnII, respectively. Residues 314–323 of 6FnI, and 414–421 and 448–449 of 2FnII are primarily involved in forming the interface, with significant contributions from Met320, Ser415, Ala418, Leu419, Thr448, and Thr449 (Fig. 2A).

Bottom Line: Analysis of NMR titrations with single-stranded collagen peptides reveals a dominant collagen interaction surface on domains (2)FnII and (7)FnI; a similar surface appears involved in interactions with triple-helical peptides.Models of the complete GBD, based on the new structure and the (8-9)FnI·collagen complex show a continuous putative collagen binding surface.We explore the implications of this model using long collagen peptides and discuss our findings in the context of FN interactions with collagen fibrils.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.

ABSTRACT
Collagen and fibronectin (FN) are two abundant and essential components of the vertebrate extracellular matrix; they interact directly with cellular receptors and affect cell adhesion and migration. Past studies identified a FN fragment comprising six modules, (6)FnI(1-2)FnII(7-9)FnI, and termed the gelatin binding domain (GBD) as responsible for collagen interaction. Recently, we showed that the GBD binds tightly to a specific site within type I collagen and determined the structure of domains (8-9)FnI in complex with a peptide from that site. Here, we present the crystallographic structure of domains (6)FnI(1-2)FnII(7)FnI, which form a compact, globular unit through interdomain interactions. Analysis of NMR titrations with single-stranded collagen peptides reveals a dominant collagen interaction surface on domains (2)FnII and (7)FnI; a similar surface appears involved in interactions with triple-helical peptides. Models of the complete GBD, based on the new structure and the (8-9)FnI·collagen complex show a continuous putative collagen binding surface. We explore the implications of this model using long collagen peptides and discuss our findings in the context of FN interactions with collagen fibrils.

Show MeSH
Related in: MedlinePlus