Ephrin-B2-induced Cleavage of EphB2 Receptor Is Mediated by Matrix Metalloproteinases to Trigger Cell Repulsion

Abstract

EphB receptors provide crucial adhesive and repulsive signals during cell migration and axon guidance, but it is unclear how they switch between these opposing responses. Here we provide evidence of an important role for matrix metalloproteinases (MMPs) in repulsive EphB2 signaling. We found that EphB2 is cleaved by MMPs both in vitro and in vivo, and that this cleavage is induced by interaction with its ligand ephrin-B2. Our findings demonstrate that MMP-2/MMP-9-specific inhibition or cleavage-resistant mutations in of EphB2 in and in of but and in inhibition of EphB2 cleavage EphB2 evidence that cleavage of EphB2 is induced by cell and that this responses. EphB receptors provide crucial adhesive and repulsive signals during cell migration and axon guidance, but it is unclear how they switch between these opposing responses. Here we provide evidence of an important role for matrix metalloproteinases (MMPs) in repulsive EphB2 signaling. We found that EphB2 is cleaved by MMPs both in vitro and in vivo, and that this cleavage is induced by interaction with its ligand ephrin-B2. Our findings demonstrate that MMP-2/MMP-9-specific inhibition or cleavage-resistant mutations in of EphB2 in and in of but and in inhibition of EphB2 cleavage EphB2 evidence that cleavage of EphB2 is induced by cell and that this responses. matrix in cell cell of matrix in cell cell of receptors of that important of cell axon guidance, and of receptors by or responses. both receptors and between receptors and is unclear how between and is that repulsive cell induced by of EphB2 cleavage by by cleavage of and by or EphB of EphB2 cell in of it cleavage of EphB receptors that cell metalloproteinases (MMPs) of metalloproteinases that matrix in of cell and cleavage of and in these important in and of MMPs and of is in and We that MMPs and in for MMPs in and is with MMPs and role in of adhesive we between EphB2 cell and of EphB2 We of MMPs and EphB2 and cleavage in EphB2 of these ligand but EphB2 and cell findings demonstrate for that cleavage an important role in repulsive of EphB2 cell EphB2 by of and or mutations of cleavage by or in and or in of EphB2 and EphB2 in in vitro cleavage of of and of with with or or in with cell or by with of EphB2 by and or of with or and with or in cleavage and for for with in and EphB2 cleavage by with of or EphB2 and with of or with for with EphB2 or with of or with or MMP-2/MMP-9-specific and and in with or for with of or by of or with for of or in and cell with for and EphB2 cleavage by an and with an of and with an EphB2 and of or with or for or in and cell with and for with with and an and with and or with or for in and of cell by and with and by with of by and in cell and in and and or of and for with and with of with and and of and or and of for by by or and and in with of an with and by in with and and of between EphB2 and for of and and and for of for for by by with in and in and in with and and or of and for and with or for and in and in by and of cell and of or of of for with and and for for by by or with or EphB2 or for for and an with or with and for for by by of or with and in and in for and for in with of or of with of and of EphB2 with and with and for with and and for and of cell of of EphB2 or and of for for by by of EphB2 EphB2 interaction with its ligand of EphB2 in EphB2 and of and in cell of an for of EphB2 of EphB2 with of in that interaction cleavage of EphB2 of in of an for of that EphB2 cleavage cell and in of but of and that these cleavage in during an of and cleavage of EphB2 in and findings that EphB2 is cleaved both in vitro and in this cleavage is induced by interaction and by and EphB2 and EphB2 in vitro and in of EphB2 and and with or an and of with an that of EphB2 and in that by of cleavage in of EphB2 EphB2 cleaved by and and of EphB2 and in EphB2 cleaved by EphB2 cleavage that in and in and in and cleavage EphB2 cleaved by in and and in in these findings demonstrate that MMPs of EphB2 in and of and EphB2 in vitro and in and of EphB2 and of with and in and in of and with of EphB2 of and cleaved by and in an with of EphB2 in with and cleavage of EphB2 in by cleavage in with EphB2 and in with cleaved by in of and in with with an of EphB2 with of and EphB2 cleavage by cleavage in and of EphB2 and in EphB2 mutations in and in and in and in EphB2 cleavage by MMPs cleavage of EphB2 mutations and of in in but in both and and is receptors is EphB2 and receptors but is found in that in cleavage of repulsive by EphB findings demonstrate that of EphB2 cleavage of cleavage in of EphB2 cleavage of EphB2 in by of cleavage in that or cleavage in in but that is by in with that EphB2 cleavage is induced with in but with with in of EphB2 cleavage by we of mutations and of EphB2 We cell that or of in of and or that induced of or and responses. of or that or but of or that adhesive we cell between ligand and EphB2 and EphB2 or with and of and of that of of but adhesive and adhesive and of we mutations for with and and with of and for demonstrate that mutations cleavage and in EphB2 ligand of but of EphB2 and in by cleavage of EphB or we and EphB We and by for that of We found of in with with with and of in of that and by of in with but with of by that this of in that it of EphB2 signaling. is that EphB2 cleavage and and in is by of in of and or or with by by of and of or or with that by these of in or in in with in and EphB2 we and in or of or with and or and or for with and of in with EphB in of or with or findings demonstrate that and by EphB2 We of cleavage in these by and in EphB or in but of in and in and in and in with findings demonstrate that cleavage of EphB2 is in and in and in EphB2 of in of and by of and and of and of or and or and in with or EphB2 repulsive with cleavage of of that cleavage of EphB2 signaling. we by its ligand and induced of both and in and of and and of and by in of with and its with with of and its with and for of inhibition of with of EphB2 and of and EphB2 its with these demonstrate that inhibition of EphB2 cleavage with or cleavage-resistant in EphB2 EphB2 inhibition of EphB2 cleavage EphB2 and of of and and of and with in by with and with or with for of or by with of EphB2 or of and with and by and EphB2 of with induced of both and and of and of and of and in of with and its with with for EphB receptors with or with for with and with of EphB2 or of and with EphB2 by and EphB2 of and EphB2 by we cleavage of EphB2 its we of in or of in but in with in but in findings demonstrate that cleavage of EphB2 its is by inhibition of EphB2 of or or with or of in but that of in by of of in of and by MMPs and EphB receptors important in and of but of Here we provide evidence for role of MMPs in repulsive EphB2 signaling. Our findings that EphB cleavage is induced by interaction and by EphB2 cleavage and cleavage-resistant mutations cleavage in of EphB2 its and that EphB2 cleavage by that of EphB2 of EphB2 that an EphB2 Our that EphB2 cleavage cell that is cleaved cleavage is by Our findings demonstrate that EphB2 is cleaved by MMPs cell in interaction with and cleavage by is for cleavage of and and by or by of by of is by an of is cleavage is or but findings evidence that is of cleavage of cleavage in of EphB2 that receptors cleavage is or and in with of is found in EphB2 and that in of EphB receptors of repulsive by EphB receptors of EphB2 cleavage in these EphB2 in and important in of EphB2 in of and that in and and it is that EphB2 cleavage an important role in these for Here we demonstrate that EphB2 is cleaved by and and this cleavage is induced by and MMPs in and during in and in in of responses. we that cleavage-resistant mutations cleavage in of EphB2 or and of of we of inhibition EphB2 Our findings that cleavage repulsive by EphB2 of cleavage-resistant mutations in EphB2 induced of its with ephrin-B2. cleavage of EphB2 of EphB2 with of EphB2 cleavage and but of and EphB2 its in repulsive and that an interaction that is by that for and both and EphB receptors and of in EphB2 and EphB2 its by with its or by is an that EphB2 and of of EphB2 an it is that of EphB2 its and we demonstrate that cleavage of EphB2 is important for EphB2 EphB2 cleavage with or cleavage-resistant mutations EphB2 of and and Our that EphB2 in of of and of that and and of cell and cell in and of and cell its of EphB receptors by or cleavage of EphB2 an important role in cell migration and axon findings demonstrate that cleavage of EphB2 is by MMPs and and matrix in cell cell of matrix in cell cell of receptors of that important of cell axon guidance, and of receptors by or responses. both receptors and between receptors and is unclear how between and is that repulsive cell induced by of EphB2 cleavage by by cleavage of and by or EphB of EphB2 cell in of it cleavage of EphB receptors that cell metalloproteinases (MMPs) of metalloproteinases that matrix in of cell and cleavage of and in these important in and of MMPs and of is in and We that MMPs and in for MMPs in and is with MMPs and role in of adhesive Here we between EphB2 cell and of EphB2 We of MMPs and EphB2 and cleavage in EphB2 of these ligand but EphB2 and cell findings demonstrate for that cleavage an important role in repulsive of EphB2 cell EphB2 by of and or mutations of cleavage by or in and or in of EphB2 and EphB2 in in vitro cleavage of of and of with with or or in with cell or by with of EphB2 by and or of with or and with or in cleavage and for for with in and EphB2 cleavage by with of or EphB2 and with of or with for with EphB2 or with of or with or MMP-2/MMP-9-specific and and in with or for with of or by of or with for of or in and cell with for and EphB2 cleavage by an and with an of and with an EphB2 and of or with or for or in and cell with and for with with and an and with and or with or for in and of cell by and with and by with of by and in cell and in and and or of and for with and with of with and and of and or and of for by by or and and in with of an with and by in with and and of between EphB2 and for of and and and for of for for by by with in and in and in with and and or of and for and with or for and in and in by and of cell and of or of of for with and and for for by by or with or EphB2 or for for and an with or with and for for by by of or with and in and in for and for in with of or of with of and of EphB2 with and with and for with and and for and of cell of of EphB2 or and of for for by by cell EphB2 by of and or mutations of cleavage by or in and or in of EphB2 and EphB2 in in vitro cleavage of of cell EphB2 by of and or mutations of cleavage by or in and or in of EphB2 and EphB2 in in vitro cleavage of of and of with with or or in with cell or by with of with with or or in with cell or by with of EphB2 by and or of with or and with or in cleavage and for for with in and EphB2 cleavage by with of or EphB2 and with of or with for with EphB2 or with of or with or MMP-2/MMP-9-specific and and in with or for with of or by of or with for of or in and cell with for and EphB2 cleavage by an and with an of and with an EphB2 and of or with or for or in and cell with and for with with and an and with and or with or for in and of cell by and with and by with of by and in cell and in and and or of and for with and with of with and and of and or and of for by by or and and in with of an with and by in with and and of between EphB2 and for of and and and for of for for by by with in and in and in with and and or of and for and with or for and in and in by and of cell and of or of of for with and and for for by by or with or EphB2 or for for and an with or with and for for by by of or with and in and in for and for in with of or of with of and of EphB2 with and with and for with and and for and of cell of of EphB2 or and of for for by by or of with or and with or in cleavage and for for with in and EphB2 cleavage by with of or EphB2 and with of or with for with EphB2 or with of or with or MMP-2/MMP-9-specific and and in with or for with of or by of or with for of or in and cell with for and EphB2 cleavage by an and with an of and with an EphB2 and of or with or for or in and cell with and for with with and an and with and or with or for in and of cell by and with and by with of by and in cell and in and and or of and for with and with of with and and of and or and of for by by or and and in with of an with and by in with and and of between EphB2 and for of and and and for of for for by by with in and in and in with and and or of and for and with or for and in and in by and of cell and of or of of for with and and for for by by or with or EphB2 or for for and an with or with and for for by by of or with and in and in for and for in with of or of with of and of EphB2 with and with and for with and and for and of cell of of EphB2 or and of for for by by of EphB2 EphB2 interaction with its ligand of EphB2 in EphB2 and of and in cell of an for of EphB2 of EphB2 with of in that interaction cleavage of EphB2 of in of an for of that EphB2 cleavage cell and in of but of and that these cleavage in during an of and cleavage of EphB2 in and findings that EphB2 is cleaved both in vitro and in this cleavage is induced by interaction and by and EphB2 and EphB2 in vitro and in of EphB2 and and with or an and of with an that of EphB2 and in that by of cleavage in of EphB2 EphB2 cleaved by and and of EphB2 and in EphB2 cleaved by EphB2 cleavage that in and in and in and cleavage EphB2 cleaved by in and and in in these findings demonstrate that MMPs of EphB2 in and of cleavage in and of EphB2 and in EphB2 mutations in and in and in and in EphB2 cleavage by MMPs cleavage of EphB2 mutations and of in in but in both and and is receptors is EphB2 and receptors but is found in that in cleavage of repulsive by EphB findings demonstrate that of EphB2 cleavage of cleavage in of EphB2 cleavage of EphB2 in by of cleavage in that or cleavage in in but that is by in with that EphB2 cleavage is induced with in but with with in of EphB2 cleavage by we of mutations and of EphB2 We cell that or of in of and or that induced of or and responses. of or that or but of or that adhesive we cell between ligand and EphB2 and EphB2 or with and of and of that of of but adhesive and adhesive and of we mutations for with and and with of and for demonstrate that mutations cleavage and in EphB2 ligand of but of EphB2 and in by cleavage of EphB or we and EphB We and by for that of We found of in with with with and of in of that and by of in with but with of by that this of in that it of EphB2 signaling. is that EphB2 cleavage and and in is by of in of and or or with by by of and of or or with that by these of in or in in with in and EphB2 we and in or of or with and or and or for with and of in with EphB in of or with or findings demonstrate that and by EphB2 We of cleavage in these by and in EphB or in but of in and in and in and in with findings demonstrate that cleavage of EphB2 is in and in and in EphB2 of in of and by of and and of and of or and or and in with or EphB2 repulsive with cleavage of of that cleavage of EphB2 signaling. we by its ligand and induced of both and in and of and and of and by in of with and its with with of and its with and for of inhibition of with of EphB2 and of and EphB2 its with these demonstrate that inhibition of EphB2 cleavage with or cleavage-resistant in EphB2 EphB2 inhibition of EphB2 cleavage EphB2 and of of and and of and with in by with and with or with for of or by with of EphB2 or of and with and by and EphB2 of with induced of both and and of and of and of and in of with and its with with for EphB receptors with or with for with and with of EphB2 or of and with EphB2 by and EphB2 of and EphB2 by we cleavage of EphB2 its we of in or of in but in with in but in findings demonstrate that cleavage of EphB2 its is by inhibition of EphB2 of or or with or of in but that of in by of of in of and by of EphB2 EphB2 interaction with its ligand of EphB2 in EphB2 and of and in cell of an for of EphB2 of EphB2 with of in that interaction cleavage of EphB2 of in of an for of that EphB2 cleavage cell and in of but of and that these cleavage in during an of and cleavage of EphB2 in and findings that EphB2 is cleaved both in vitro and in this cleavage is induced by interaction and by and EphB2 and EphB2 in vitro and in of EphB2 and and with or an and of with an that of EphB2 and in that by of cleavage in of EphB2 EphB2 cleaved by and and of EphB2 and in EphB2 cleaved by EphB2 cleavage that in and in and in and cleavage EphB2 cleaved by in and and in in these findings demonstrate that MMPs of EphB2 in and of cleavage in and of EphB2 and in EphB2 mutations in and in and in and in EphB2 cleavage by MMPs cleavage of EphB2 mutations and of in in but in both and and is receptors is EphB2 and receptors but is found in that in cleavage of repulsive by EphB findings demonstrate that of EphB2 cleavage with in of EphB2 cleavage by we of mutations and of EphB2 We cell that or of in of and or that induced of or and we cell between ligand and EphB2 and EphB2 or with and of and of that of of but adhesive and adhesive and of we mutations for with and and with of and for demonstrate that mutations cleavage and in EphB2 ligand of but of EphB2 and in by cleavage of EphB or we and EphB We and by for that of We found of in with with with and of in of that and by of in with but with of by that this of in that it of EphB2 signaling. is that EphB2 cleavage and EphB2 we and in or of or with and or and or for with and of in with EphB in of or with or findings demonstrate that and by EphB2 We of cleavage in these by and in EphB or in but of in and in and in and in with findings demonstrate that cleavage of EphB2 is in and in EphB2 repulsive with cleavage of of that cleavage of EphB2 signaling. we by its ligand and induced of both and in and of and and of and by in of with and its with with of and its with and for of inhibition of with of EphB2 and of and EphB2 its with these demonstrate that inhibition of EphB2 cleavage with or cleavage-resistant in EphB2 EphB2 by we cleavage of EphB2 its we of in or of in but in with in but in findings demonstrate that cleavage of EphB2 its MMPs and EphB receptors important in and of but of Here we provide evidence for role of MMPs in repulsive EphB2 signaling. Our findings that EphB cleavage is induced by interaction and by EphB2 cleavage and cleavage-resistant mutations cleavage in of EphB2 its and that EphB2 cleavage by that of EphB2 of EphB2 that an EphB2 Our that EphB2 cleavage cell that is cleaved cleavage is by Our findings demonstrate that EphB2 is cleaved by MMPs cell in interaction with and cleavage by is for cleavage of and and by or by of by of is by an of is cleavage is or but findings evidence that is of cleavage of cleavage in of EphB2 that receptors cleavage is or and in with of is found in EphB2 and that in of EphB receptors of repulsive by EphB receptors of EphB2 cleavage in these EphB2 in and important in of EphB2 in of and that in and and it is that EphB2 cleavage an important role in these for Here we demonstrate that EphB2 is cleaved by and and this cleavage is induced by and MMPs in and during in and in in of responses. we that cleavage-resistant mutations cleavage in of EphB2 or and of of we of inhibition EphB2 Our findings that cleavage repulsive by EphB2 of cleavage-resistant mutations in EphB2 induced of its with ephrin-B2. cleavage of EphB2 of EphB2 with of EphB2 cleavage and but of and EphB2 its in repulsive and that an interaction that is by that for and both and EphB receptors and of in EphB2 and EphB2 its by with its or by is an that EphB2 and of of EphB2 an it is that of EphB2 its and we demonstrate that cleavage of EphB2 is important for EphB2 EphB2 cleavage with or cleavage-resistant mutations EphB2 of and and Our that EphB2 in of of and of that and and of cell and cell in and of and cell its of EphB receptors by or cleavage of EphB2 an important role in cell migration and axon findings demonstrate that cleavage of EphB2 is by MMPs and and MMPs and EphB receptors important in and of but of Here we provide evidence for role of MMPs in repulsive EphB2 signaling. Our findings that EphB cleavage is induced by interaction and by EphB2 cleavage and cleavage-resistant mutations cleavage in of EphB2 its and that EphB2 cleavage by that of EphB2 of EphB2 that an EphB2 Our that EphB2 cleavage cell that is cleaved cleavage is by Our findings demonstrate that EphB2 is cleaved by MMPs cell in interaction with and cleavage by is for cleavage of and and by or by of by of is by an of is cleavage is or but findings evidence that is of cleavage Our of cleavage in of EphB2 that receptors cleavage is or and in with of is found in EphB2 and that in of EphB receptors of repulsive by EphB receptors of EphB2 cleavage in these EphB2 in and important in of EphB2 in of and that in and and it is that EphB2 cleavage an important role in these responses. for Here we demonstrate that EphB2 is cleaved by and and this cleavage is induced by and MMPs in and during in and in in of responses. we that cleavage-resistant mutations cleavage in of EphB2 or and of of we of inhibition EphB2 Our findings that cleavage repulsive by EphB2 of cleavage-resistant mutations in EphB2 induced of its with ephrin-B2. cleavage of EphB2 of EphB2 with of EphB2 cleavage and but of and EphB2 its in repulsive and that an interaction that is by that for and both and EphB receptors and of in EphB2 and responses. EphB2 its by with its or by is an that EphB2 and of of EphB2 an it is that of EphB2 its and Here we demonstrate that cleavage of EphB2 is important for EphB2 EphB2 cleavage with or cleavage-resistant mutations EphB2 of and and Our that EphB2 in of of and of that and and of cell and cell in and of and cell its of EphB receptors by or cleavage of EphB2 an important role in cell migration and axon findings demonstrate that cleavage of EphB2 is by MMPs and and We for and for We of for with with