A Peptide Derived from Tenascin-C Induces β1 Integrin Activation through Syndecan-4

Tenascin-C (TN-C) is unique for its cell adhesion modulatory function. We have shown that TNIIIA2, a synthetic 22-mer peptide derived from TN-C, stimulated β1 integrin-mediated cell adhesion of nonadherent and adherent cell types, by inducing activation of β1 integrin. The active site of TNIIIA2 appeared cryptic in the TN-C molecule but was exposed by MMP-2 processing of TN-C. The following results suggest that cell surface heparan sulfate (HS) proteoglycan (HSPG), including syndecan-4, participated in TNIIIA2-induced β1 integrin activation: 1) TNIIIA2 bound cell surface its by of β1 integrin activation by was by of TNIIIA2-induced β1 integrin and cell adhesion of TNIIIA2-induced activation of β1 integrin. that the of syndecan-4, including of its the and the TNIIIA2 results suggest that a site of the TN-C exposed by MMP-2 have bound its and in β1 integrin for its of β1 integrin the of the molecule in Tenascin-C (TN-C) is unique for its cell adhesion modulatory function. We have shown that TNIIIA2, a synthetic 22-mer peptide derived from TN-C, stimulated β1 integrin-mediated cell adhesion of nonadherent and adherent cell types, by inducing activation of β1 integrin. The active site of TNIIIA2 appeared cryptic in the TN-C molecule but was exposed by MMP-2 processing of TN-C. The following results suggest that cell surface heparan sulfate (HS) proteoglycan (HSPG), including syndecan-4, participated in TNIIIA2-induced β1 integrin activation: 1) TNIIIA2 bound cell surface its by of β1 integrin activation by was by of TNIIIA2-induced β1 integrin and cell adhesion of TNIIIA2-induced activation of β1 integrin. that the of syndecan-4, including of its the and the TNIIIA2 results suggest that a site of the TN-C exposed by MMP-2 have bound its and in β1 integrin for its of β1 integrin the of the molecule in TN-C, heparan sulfate of TN-C, heparan sulfate of is of the TN-C is and in the TN-C have for TN-C, including cell and The of TN-C cell adhesion the TN-C of cell types, but is for cell cell TN-C is and is adhesion modulatory a TN-C molecule including the have in its a the adhesion modulatory of TN-C of the by of the integrin of The unique of is and integrin-mediated in in for the and activation is by from the cell that by the activation of have that and have active cryptic exposed and in of the active of and of have in a of by have that cell adhesion and a site cell The 22-mer derived from the β1 integrin-mediated cell adhesion its cryptic site is exposed by a of and the of in including TN-C. and in the of the and the of the TN-C molecule in TN-C cell adhesion modulatory We have the of synthetic TN-C cell adhesion TNIIIA2, a 22-mer TN-C peptide stimulated cell adhesion by inducing and activation of β1 integrin. The active site of TNIIIA2 cryptic but was exposed by MMP-2 of results suggest that cell surface heparan sulfate (HS) (HSPG), including syndecan-4, participated in β1 integrin activation in TNIIIA2, its a site of TN-C exposed by MMP-2 a of β1 integrin the in in β1 integrin for its The cell of TN-C, in the and and in and and in and of and of for for by following and in of by the was by was from derived from TN-C and and the the and in a was the of peptide by and a of a cell surface of TNIIIA2, a of TNIIIA2, shown the TNIIIA2 was synthetic by and by integrin and from that integrin β1 was from and that integrin β1 was from the active of integrin and the heparan sulfate (HS) and a by of from and integrin from was from TNIIIA2 for and of β1 the The and β1 and and a and adhesion was cell adhesion was of β1 of β1 the was by and and the active β1 integrin TN-C and the of TN-C was the was by the and of and was in a was for and the MMP-2 and was the for the MMP-2 and the a was the a for in the of and for a of of and of the of the was the the the the bound and and and was for have the the the bound the was and and for and and and by of and the from The by and the for of was by and of by a synthetic TN-C and in TNIIIA2 and cell adhesion of the synthetic the of the the in the of TNIIIA2, and the was a of and in the is and and cell adhesion in a and was by integrin and but and of TNIIIA2 integrin-mediated cell and and and in in the of TNIIIA2 peptide a the in and in and a was a and for for and the of was the of of is the of TNIIIA2 adhesion of nonadherent cell a β1 and of the β1 in in the of integrin TNIIIA2 was of inducing cell and was by and but TNIIIA2 cell and was by and but by for and and TNIIIA2 cell in the of TNIIIA2 nonadherent cell adhesion that peptide activation of β1 of β1 of β1 integrin activation by its integrin and of the active of β1 of the We TNIIIA2 was the of β1 integrin and activation of β1 in TNIIIA2 TNIIIA2 for the integrin shown of and of active β1 cell for in the of TNIIIA2 and results in of the of a of integrin a in of the and the TNIIIA2 a in of the in a and of TNIIIA2 a but in of the TNIIIA2 of the β1 integrin the by and of the active in TNIIIA2, was by nonadherent cell in the β1 integrin was adherent cell The by a in TNIIIA2, the nonadherent cell types, by and in nonadherent and adherent cell types, of the and in TNIIIA2 the of peptide cell adhesion that TNIIIA2 stimulated β1 integrin-mediated cell adhesion by inducing β1 integrin of β1 by the and a of β1 the the stimulated TNIIIA2, the of appeared the a in the of the the of the was the but TNIIIA2 The results that TNIIIA2 a in the of β1 integrin cell of TNIIIA2 in the TN-C the active site of TNIIIA2 in the TN-C the in the was for the the and the results of β1 integrin by The TNIIIA2 was of the and and and the for β1 integrin The of in TNIIIA2 of in of the the of a and for β1 integrin activation in the active site of TNIIIA2 is exposed the TN-C the of a of the cell the cell was by processing of but a in cell The MMP-2 of β1 integrin activation the active site the TN-C but exposed by processing of the site by processing of was MMP-2 and for cell adhesion of β1 integrin activation of TNIIIA2 and MMP-2 cell adhesion in the of of is of MMP-2 β1 integrin activation in in results in of in of TNIIIA2 the of a TNIIIA2 a cell surface TNIIIA2, by We a molecule by peptide its a the molecule by a TNIIIA2 the of a and of and the from the and in the of The was and by was by and the was in the of the of and TNIIIA2 was shown sulfate and β1 integrin activation in TNIIIA2 was in the of results suggest that TNIIIA2 its cell surface its β1 integrin was but a a from that of and of β1 integrin cell surface in and of a cell surface molecule TNIIIA2 by and of 1) and was in in the and of a the and integrin and by the was 1) of the and was in in shown of the was of the the a was and its a of and including a the The of was by of and but the cell surface have the in TNIIIA2-induced β1 integrin of of the of in the of TNIIIA2 the of TNIIIA2-induced β1 integrin of β1 integrin activation in TNIIIA2 We the of by and that was by of TNIIIA2 β1 integrin activation and cell adhesion in a of that activation a in cell results suggest that the and its for β1 integrin activation by in TNIIIA2-induced β1 integrin of for TNIIIA2-induced β1 integrin for TNIIIA2 and for β1 integrin activation in is in of of of TNIIIA2 and in by and the and and The is by results in and a and and and a and TNIIIA2 and for β1 integrin activation cell adhesion results in of and its in the for and We its of the of a in β1 integrin activation by TNIIIA2 of but β1 integrin activation in TNIIIA2 the a TNIIIA2-induced β1 integrin activation results suggest that activation of β1 integrin by TNIIIA2 cell surface including syndecan-4, but of its β1 integrin activation of and and the was by TNIIIA2 and for β1 integrin activation in active of β1 integrin in a for and in the of TNIIIA2 β1 integrin activation was by in is in and results in in a in and of of the TN-C molecule that and TN-C. a TN-C a of TN-C, cell the site in of TN-C the The and that TN-C cell of of that TN-C cell by the of the of syndecan-4, and activation of and TN-C and and in TN-C a for cell adhesion and by of cell adhesion by integrin and the results of the that the TN-C molecule a site that cell adhesion synthetic TN-C TNIIIA2, the activation of β1 by active β1 and We have that TNIIIA2 from by the is of integrin in cell The of its integrin function. that in β1 integrin in TNIIIA2 its that integrin activation is by from the cell is by in β1 integrin and and integrin including and the of β1 integrin. is that the of β1 integrin in β1 integrin surface integrin and integrin the the and of the that a for a integrin. that the a the of and the of the of in cell adhesion a of have shown the of the and site the of surface molecule adhesion the The integrin was shown for activation and for cell and have shown that a is for cell adhesion of the have shown that cell adhesion the β1 that in β1 integrin activation by TNIIIA2-induced β1 integrin activation the the of the TNIIIA2-induced β1 integrin activation a the of and β1 integrin in β1 integrin. the a of β1 integrin cell surface that a the β1 integrin its a active the active of β1 integrin TNIIIA2 a in the of β1 cell in adhesion of adherent and nonadherent cell adherent cell types, the and in and TNIIIA2 a is TNIIIA2 adhesion of nonadherent cell types, in that of adherent cell of TN-C in and of TN-C in and is a of TN-C in the a in and in integrin the a for β1 integrin activation a site the TN-C is TNIIIA2 in β1 integrin for its that β1 integrin is its syndecan-4, the of cell adhesion the of in adhesion and in that the active site of TNIIIA2 cryptic in the TN-C that TNIIIA2 in in of a of for cell have shown that of derived from active cryptic of the site was exposed by have shown that of TN-C and that MMP-2 a the that TN-C, its the is in and in of the and of of is the in TN-C and syndecan-4, is that the in syndecan-4, a for the of was that in the the site is the of TNIIIA2 for and the of the is the site is exposed its in that a site cell TNIIIA2, peptide site is of β1 is that the a site that β1 integrin-mediated cell adhesion and the TN-C a site that that and TN-C active that in of and TN-C a for TN-C, heparan sulfate of TN-C, heparan sulfate of is of the TN-C is and in the TN-C have for TN-C, including cell and The of TN-C cell adhesion the TN-C of cell types, but is for cell cell TN-C is and is adhesion modulatory a TN-C molecule including the have in its a the adhesion modulatory of TN-C of the by of the integrin of The unique of is and integrin-mediated in in for the and activation is by from the cell that by the activation of have that and have active cryptic exposed and in of the active of and of have in a of by We have that cell adhesion and a site cell The 22-mer derived from the β1 integrin-mediated cell adhesion its cryptic site is exposed by a of and the of in including TN-C. and in the of the and the of the TN-C molecule in TN-C cell adhesion modulatory We have the of synthetic TN-C cell adhesion TNIIIA2, a 22-mer TN-C peptide stimulated cell adhesion by inducing and activation of β1 integrin. The active site of TNIIIA2 cryptic but was exposed by MMP-2 of results suggest that cell surface heparan sulfate (HS) (HSPG), including syndecan-4, participated in β1 integrin activation in TNIIIA2, its a site of TN-C exposed by MMP-2 a of β1 integrin the in in β1 integrin for its The cell of TN-C, in the and and in and and in and of and of for for by following and in of by the was by was from derived from TN-C and and the the and in a was the of peptide by and a of a cell surface of TNIIIA2, a of TNIIIA2, shown the TNIIIA2 was synthetic by and by integrin and from that integrin β1 was from and that integrin β1 was from the active of integrin and the heparan sulfate (HS) and a by of from and integrin from was from TNIIIA2 for and of β1 the The and β1 and and a and adhesion was cell adhesion was of β1 of β1 the was by and and the active β1 integrin TN-C and the of TN-C was the was by the and of and was in a was for and the MMP-2 and was the for the MMP-2 and the a was the a for in the of and for a of of and of the of the was the the the the bound and and and was for have the the the bound the was and and for and and and by of and the from The by and the for of was by and and and in and and in and of and of for for by following and in of by the was by was from derived from TN-C and and the the and in a was the of peptide by and a of a cell surface of TNIIIA2, a of TNIIIA2, shown the TNIIIA2 was synthetic by and by integrin and from that integrin β1 was from and that integrin β1 was from the active of integrin and the heparan sulfate (HS) and a by of from and integrin from was from TNIIIA2 for and of β1 the The and β1 and and a and adhesion was cell adhesion was of β1 of β1 the was by and and the active β1 integrin TN-C and the of TN-C was the was by the and of and was in a was for and the MMP-2 and was the for the MMP-2 and the a was the a for in the of and for a of of and of the of the was the the the the bound and and and was for have the the the bound the was and and for and and and by of and the from The by and the for of was by and of by a synthetic TN-C and in TNIIIA2 and cell adhesion of the synthetic the of the the in the of TNIIIA2, and the was a of and in the is and and cell adhesion in a and was by integrin and but and the of TNIIIA2 adhesion of nonadherent cell a β1 and of the β1 in in the of integrin TNIIIA2 was of inducing cell and was by and but TNIIIA2 cell and was by and but by for and and TNIIIA2 cell in the of TNIIIA2 nonadherent cell adhesion that peptide activation of β1 of β1 of β1 integrin activation by its integrin and of the active of β1 of the We TNIIIA2 was the of β1 integrin and activation of β1 in TNIIIA2 TNIIIA2 for the integrin shown of and of active β1 cell for in the of TNIIIA2 and results in of the of a of integrin a in of the and the TNIIIA2 a in of the in a and of TNIIIA2 a but in of the TNIIIA2 of the β1 integrin the by and of the active in TNIIIA2, was by nonadherent cell in the β1 integrin was adherent cell The by a in TNIIIA2, the nonadherent cell types, by and in nonadherent and adherent cell types, of the and in TNIIIA2 the of peptide cell adhesion that TNIIIA2 stimulated β1 integrin-mediated cell adhesion by inducing β1 integrin of β1 by the and a of β1 the the stimulated TNIIIA2, the of appeared the a in the of the the of the was the but TNIIIA2 The results that TNIIIA2 a in the of β1 integrin cell of TNIIIA2 in the TN-C the active site of TNIIIA2 in the TN-C the in the was for the the and the results of β1 integrin by The TNIIIA2 was of the and and and the for β1 integrin The of in TNIIIA2 of in of the the of a and for β1 integrin activation in the active site of TNIIIA2 is exposed the TN-C the of a of the cell the cell was by processing of but a in cell The MMP-2 of β1 integrin activation the active site the TN-C but exposed by processing of the site by processing of was MMP-2 and for cell adhesion of β1 integrin activation of TNIIIA2 and MMP-2 cell adhesion in the of of is of MMP-2 β1 integrin activation in in results in of in of TNIIIA2 the of a TNIIIA2 a cell surface TNIIIA2, by We a molecule by peptide its a the molecule by a TNIIIA2 the of a and of and the from the and in the of The was and by was by and the was in the of the of and TNIIIA2 was shown sulfate and β1 integrin activation in TNIIIA2 was in the of results suggest that TNIIIA2 its cell surface its β1 integrin was but a a from that of and of β1 integrin cell surface in and of a cell surface molecule TNIIIA2 by and of 1) and was in in the and of a the and integrin and by the was 1) of the and was in in shown of the was of the the a was and its a of and including a the The of was by of and but the cell surface have the in TNIIIA2-induced β1 integrin of of the of in the of TNIIIA2 the of TNIIIA2-induced β1 integrin of β1 integrin activation in TNIIIA2 We the of by and that was by of TNIIIA2 β1 integrin activation and cell adhesion in a of that activation a in cell results suggest that the and its for β1 integrin activation by in TNIIIA2-induced β1 integrin of for TNIIIA2-induced β1 integrin for TNIIIA2 and for β1 integrin activation in is in of of of TNIIIA2 and in by and the and and The is by results in and a and and and a and TNIIIA2 and for β1 integrin activation cell adhesion results in of and its in the for and We its of the of a in β1 integrin activation by TNIIIA2 of but β1 integrin activation in TNIIIA2 the a TNIIIA2-induced β1 integrin activation results suggest that activation of β1 integrin by TNIIIA2 cell surface including syndecan-4, but of its β1 integrin activation of and and the was by TNIIIA2 and for β1 integrin activation in active of β1 integrin in a for and in the of TNIIIA2 β1 integrin activation was by in is in and results in of by a synthetic TN-C and in TNIIIA2 and cell adhesion of the synthetic the of the the in the of TNIIIA2, and the was a of and in the is and and cell adhesion in a and was by integrin and but and We the of TNIIIA2 adhesion of nonadherent cell a β1 and of the β1 in in the of integrin TNIIIA2 was of inducing cell and was by and but TNIIIA2 cell and was by and but by for and and TNIIIA2 cell in the of TNIIIA2 nonadherent cell adhesion that peptide activation of β1 integrin. TNIIIA2 of β1 of β1 integrin activation by its integrin and of the active of β1 of the We TNIIIA2 was the of β1 integrin and of the of a of integrin a in of the and the TNIIIA2 a in of the in a and of TNIIIA2 a but in of the TNIIIA2 of the β1 integrin the by and of the active in TNIIIA2, was by nonadherent cell in the β1 integrin was adherent cell The by a in TNIIIA2, the nonadherent cell types, by and in nonadherent and adherent cell types, of the and in TNIIIA2 the of peptide cell adhesion that TNIIIA2 stimulated β1 integrin-mediated cell adhesion by inducing β1 integrin We of β1 by the and a of β1 the the stimulated TNIIIA2, the of appeared the a in the of the the of the was the but TNIIIA2 The results that TNIIIA2 a in the of β1 integrin cell of TNIIIA2 in the TN-C the active site of TNIIIA2 in the TN-C the in the was for the the and the results of β1 integrin by The TNIIIA2 was of the and and and the for β1 integrin The of in TNIIIA2 of in of the the of a and for β1 integrin activation in the active site of TNIIIA2 is exposed the TN-C the of a of the cell the cell was by processing of but a in cell The MMP-2 of β1 integrin activation the active site the TN-C but exposed by processing of in of TNIIIA2 the of a TNIIIA2 a cell surface TNIIIA2, by We a molecule by peptide its a the molecule by a TNIIIA2 the of a and of and the from the and in the of The was and by was by and the was in the of the of and TNIIIA2 was shown sulfate and β1 integrin activation in TNIIIA2 was in the of results suggest that TNIIIA2 its cell surface its β1 integrin was but a a from that of and of β1 integrin cell surface in the was of the the a was and its a of and including a the The of was by of and but the cell surface have the in TNIIIA2-induced β1 integrin of of the of in the of TNIIIA2 the of TNIIIA2-induced β1 integrin of β1 integrin activation in TNIIIA2 We the of by and that was by of TNIIIA2 β1 integrin activation and cell adhesion in a of that activation a in cell results suggest that the and its for β1 integrin activation by The of and its in the for and We its of the of a in β1 integrin activation by TNIIIA2 of but β1 integrin activation in TNIIIA2 the a TNIIIA2-induced β1 integrin activation results suggest that activation of β1 integrin by TNIIIA2 cell surface including syndecan-4, but of its in a in and of of the TN-C molecule that and TN-C. a TN-C a of TN-C, cell the site in of TN-C the The and that TN-C cell of of that TN-C cell by the of the of syndecan-4, and activation of and TN-C and and in TN-C a for cell adhesion and by of cell adhesion by integrin and the results of the that the TN-C molecule a site that cell adhesion synthetic TN-C TNIIIA2, the activation of β1 by active β1 and We have that TNIIIA2 from by the is of integrin in cell The of its integrin function. that in β1 integrin in TNIIIA2 its that integrin activation is by from the cell is by in β1 integrin and and integrin including and the of β1 integrin. is that the of β1 integrin in β1 integrin surface integrin and integrin the the and of the that a for a integrin. that the a the of and the of the of in cell adhesion a of have shown the of the and site the of surface molecule adhesion the The integrin was shown for activation and for cell and have shown that a is for cell adhesion of the have shown that cell adhesion the β1 that in β1 integrin activation by TNIIIA2-induced β1 integrin activation the the of the TNIIIA2-induced β1 integrin activation a the of and β1 integrin in β1 integrin. the a of β1 integrin cell surface that a the β1 integrin its a active the active of β1 integrin TNIIIA2 a in the of β1 cell in adhesion of adherent and nonadherent cell adherent cell types, the and in and TNIIIA2 a is TNIIIA2 adhesion of nonadherent cell types, in that of adherent cell of TN-C in and of TN-C in and is a of TN-C in the a in and in integrin the a for β1 integrin activation a site the TN-C is TNIIIA2 in β1 integrin for its that β1 integrin is its syndecan-4, the of cell adhesion the of in adhesion and in that the active site of TNIIIA2 cryptic in the TN-C that TNIIIA2 in in of a of for cell have shown that of derived from active cryptic of the site was exposed by have shown that of TN-C and that MMP-2 a the that TN-C, its the is in and in of the and of of is the in TN-C and syndecan-4, is that the in syndecan-4, a for the of was that in the the site is the of TNIIIA2 for and the of the is the site is exposed its in that a site cell TNIIIA2, peptide site is of β1 is that the a site that β1 integrin-mediated cell adhesion and the TN-C a site that that and TN-C active that in of and TN-C a for TN-C in a in and of of the TN-C molecule that and TN-C. a TN-C a of TN-C, cell the site in of TN-C the The and that TN-C cell of of that TN-C cell by the of the of syndecan-4, and activation of and TN-C and and in TN-C a for cell adhesion and by of cell adhesion by integrin and the results of the that the TN-C molecule a site that cell adhesion synthetic TN-C TNIIIA2, the activation of β1 by active β1 and We have that TNIIIA2 from by the is of integrin in cell The of its integrin function. that in β1 integrin in TNIIIA2 its that integrin activation is by from the cell is by in β1 integrin and and integrin including and the of β1 integrin. is that the of β1 integrin in β1 integrin surface integrin and integrin the the and of the that a for a integrin. that the a the of and the The of the of in cell adhesion a of have shown the of the and site the of surface molecule adhesion the The integrin was shown for activation and for cell and have shown that a is for cell adhesion of the have shown that cell adhesion the β1 integrin. The that in β1 integrin activation by TNIIIA2-induced β1 integrin activation the the of the TNIIIA2-induced β1 integrin activation a the of and β1 integrin in β1 integrin. the a of β1 integrin cell surface that a the β1 integrin its a active the active of β1 integrin TNIIIA2 a in the of β1 cell in adhesion of adherent and nonadherent cell adherent cell types, the and in and TNIIIA2 a is TNIIIA2 adhesion of nonadherent cell types, in that of adherent cell of TN-C in and of TN-C in and is a of TN-C in the a in and in integrin the a for β1 integrin activation a site the TN-C is TNIIIA2 in β1 integrin for its that β1 integrin is its syndecan-4, the of cell adhesion the of in adhesion and in that the active site of TNIIIA2 cryptic in the TN-C that TNIIIA2 in in of The a of for cell have shown that of derived from active cryptic of the site was exposed by have shown that of TN-C and that MMP-2 a the that TN-C, its the is in and in of the and of of is the in TN-C and syndecan-4, is that the in syndecan-4, a for the of was that in the the site is the of TNIIIA2 for and the of the is the site is exposed its in We that a site cell TNIIIA2, peptide site is of β1 is that the a site that β1 integrin-mediated cell adhesion and the TN-C a site that that and TN-C active that in of and TN-C a for