The Streptococcal Binding Site in the Gelatin-binding Domain of Fibronectin Is Consistent with a Non-linear Arrangement of Modules

TY - JOUR

T1 - The Streptococcal Binding Site in the Gelatin-binding Domain of Fibronectin Is Consistent with a Non-linear Arrangement of Modules

AU - Atkin, Kate E

AU - Brentnall, Andrew S

AU - Harris, Gemma

AU - Bingham, Richard J

AU - Erat, Michele C

AU - Millard, Christopher J

AU - Schwarz-Linek, Ulrich

AU - Staunton, David

AU - Vakonakis, Ioannis

AU - Campbell, Iain D

AU - Potts, Jennifer R

PY - 2010/11/19

Y1 - 2010/11/19

N2 - Fibronectin-binding proteins (FnBPs) of Staphylococcus aureus and Streptococcus pyogenes mediate invasion of human endothelial and epithelial cells in a process likely to aid the persistence and/or dissemination of infection. In addition to binding sites for the N-terminal domain (NTD) of fibronectin (Fn), a number of streptococcal FnBPs also contain an upstream region (UR) that is closely associated with an NTD-binding region; UR binds to the adjacent gelatin-binding domain (GBD) of Fn. Previously, UR was shown to be required for efficient streptococcal invasion of epithelial cells. Here we show, using a Streptococcus zooepidemicus FnBP, that the UR-binding site in GBD resides largely in the (8)F1(9)F1 module pair. We also show that UR inhibits binding of a peptide from the alpha 1 chain of type I collagen to (8)F1(9)F1 and that UR binding to (8)F1 is likely to occur through anti-parallel beta-zipper formation. Thus, we propose that streptococcal proteins that contain adjacent NTD- and GBD-binding sites form a highly unusual extended tandem beta-zipper that spans the two domains and mediates high affinity binding to Fn through a large intermolecular interface. The proximity of the UR- and NTD-binding sequences in streptococcal FnBPs is consistent with a non-linear arrangement of modules in the tertiary structure of the GBD of Fn.

AB - Fibronectin-binding proteins (FnBPs) of Staphylococcus aureus and Streptococcus pyogenes mediate invasion of human endothelial and epithelial cells in a process likely to aid the persistence and/or dissemination of infection. In addition to binding sites for the N-terminal domain (NTD) of fibronectin (Fn), a number of streptococcal FnBPs also contain an upstream region (UR) that is closely associated with an NTD-binding region; UR binds to the adjacent gelatin-binding domain (GBD) of Fn. Previously, UR was shown to be required for efficient streptococcal invasion of epithelial cells. Here we show, using a Streptococcus zooepidemicus FnBP, that the UR-binding site in GBD resides largely in the (8)F1(9)F1 module pair. We also show that UR inhibits binding of a peptide from the alpha 1 chain of type I collagen to (8)F1(9)F1 and that UR binding to (8)F1 is likely to occur through anti-parallel beta-zipper formation. Thus, we propose that streptococcal proteins that contain adjacent NTD- and GBD-binding sites form a highly unusual extended tandem beta-zipper that spans the two domains and mediates high affinity binding to Fn through a large intermolecular interface. The proximity of the UR- and NTD-binding sequences in streptococcal FnBPs is consistent with a non-linear arrangement of modules in the tertiary structure of the GBD of Fn.

KW - TANDEM BETA-ZIPPER

KW - STAPHYLOCOCCUS-AUREUS

KW - EPITHELIAL-CELLS

KW - MEDIATE ADHERENCE

KW - ENDOTHELIAL-CELLS

KW - NMR-SPECTROSCOPY

KW - MATRIX TURNOVER

KW - PROTEIN

KW - INTERNALIZATION

KW - BACTERIA

UR - http://www.scopus.com/inward/record.url?scp=78449251740&partnerID=8YFLogxK

U2 - 10.1074/jbc.M110.156935

DO - 10.1074/jbc.M110.156935

M3 - Article

C2 - 20843804

SN - 0021-9258

VL - 285

SP - 36977

EP - 36983

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 47

ER -