Fibulin-1 Acts as a Cofactor for the Matrix Metalloprotease ADAMTS-1

Abstract

ADAMTS-1 is a metalloprotease that has been implicated in the inhibition of angiogenesis and is a mediator of proteolytic cleavage of the hyaluronan binding proteoglycans, aggrecan and versican. In an attempt to further understand the biological function of ADAMTS-1, a yeast two-hybrid screen was performed using the carboxyl-terminal region of ADAMTS-1 as bait. As a result, the extracellular matrix protein fibulin-1 was identified as a potential interacting molecule. Through a series of analyses that included ligand affinity chromatography, co-immunoprecipitation, pulldown assays, and enzyme-linked immunosorbent assay, the ability of these two proteins to interact was substantiated. Additional studies showed that ADAMTS-1 and fibulin-1 colocalized in vivo. Furthermore, fibulin-1 was found to enhance the capacity of ADAMTS-1 to cleave aggrecan, a proteoglycan known to bind to fibulin-1. We confirmed that fibulin-1 was not a proteolytic substrate these that fibulin-1 is a of proteoglycan and an in proteoglycan in is ADAMTS-1 is a metalloprotease that has been implicated in the inhibition of angiogenesis and is a mediator of proteolytic cleavage of the hyaluronan binding proteoglycans, aggrecan and versican. In an attempt to further understand the biological function of ADAMTS-1, a yeast two-hybrid screen was performed using the carboxyl-terminal region of ADAMTS-1 as bait. As a result, the extracellular matrix protein fibulin-1 was identified as a potential interacting molecule. Through a series of analyses that included ligand affinity chromatography, co-immunoprecipitation, pulldown assays, and enzyme-linked immunosorbent assay, the ability of these two proteins to interact was substantiated. Additional studies showed that ADAMTS-1 and fibulin-1 colocalized in vivo. Furthermore, fibulin-1 was found to enhance the capacity of ADAMTS-1 to cleave aggrecan, a proteoglycan known to bind to fibulin-1. We confirmed that fibulin-1 was not a proteolytic substrate these that fibulin-1 is a of proteoglycan and an in proteoglycan in is the extracellular matrix extracellular a and metalloprotease extracellular a and metalloprotease of and as and angiogenesis ADAMTS-1 and metalloprotease to a of in the of proteins as and and proteoglycans, aggrecan, and of that ADAMTS-1 and that and In ADAMTS-1 of in the and of in the of ADAMTS-1 and to a of the of ADAMTS-1 biological and of a of that of matrix and of the the of a series of a fibulin-1 to and and to the proteoglycans, aggrecan and fibulin-1 and in the in to in to in a series of found that fibulin-1 to ADAMTS-1 and that fibulin-1 bind to aggrecan, to that fibulin-1 to enhance a was the of ADAMTS-1 was using the and and the and a of proteins was and and and and using a using and and the to in and ADAMTS-1 in a of and to proteins using and to and and of in the using a was protein to of the to fibulin-1 ADAMTS-1 using protein protein and the and proteins using and to and and in the of was and protein was to and protein and the using and ADAMTS-1 and protein and proteins using and to and of ADAMTS-1 and fibulin-1 was and ADAMTS-1 fibulin-1 in binding was binding in ADAMTS-1 fibulin-1 in and and proteins ADAMTS-1 fibulin-1 and performed using substrate and using a ADAMTS-1, in and in and and of fibulin-1 to in and and and in and and and in and and was performed using substrate of was performed using was to of the was in of and using a was and was a of in and a of in a of was of and of proteins a and of and of proteins and to of ADAMTS-1, and in in in and and and in in in in to in to in and to in was using a was in using ADAMTS-1 protein as was and ADAMTS-1 of was ADAMTS-1 and in in a of in using and to the the of was the fibulin-1 was in to fibulin-1 was of ADAMTS-1, ADAMTS-1 in in a of fibulin-1 was in fibulin-1 was of ADAMTS-1 in in a of proteins aggrecan was a is further to the and and of to in the of ADAMTS-1 was in of in of to in and aggrecan was ADAMTS-1 in the of of fibulin-1 as of aggrecan cleavage was using aggrecan ADAMTS-1 was in was and fibulin-1 was and was and of was performed as of was performed and was included a ADAMTS-1 and a the of of ADAMTS-1 of was the region to and a to the of and and was a the to of a and the and a was a was and to the to the and in and identified two and was of ADAMTS-1 was of using a was using that to the and the of was using to the of as and the was performed using of of as an yeast two-hybrid screen of a was to proteins of interacting of the carboxyl-terminal of ADAMTS-1 the two was as a to screen potential binding proteins a that to of a to the of the two and the carboxyl-terminal of the ADAMTS-1 and fibulin-1 was further to fibulin-1 to bind to ADAMTS-1 in not was to ADAMTS-1 of ADAMTS-1 in the of In the was enzyme-linked immunosorbent using As in fibulin-1 to was and ADAMTS-1 was found to bind to fibulin-1 binding affinity these two proteins was was as a has been to interact fibulin-1 ADAMTS-1, and to bind fibulin-1 a of the to a of the binding fibulin-1 to ADAMTS-1 an of as of fibulin-1 binding to As binding was not fibulin-1 and to the that fibulin-1 binding of fibulin-1 to the ADAMTS-1, fibulin-1 binding of of ADAMTS-1 to fibulin-1 of ADAMTS-1 using of of was enzyme-linked immunosorbent in the series the binding to ADAMTS-1 to fibulin-1 and the series the binding of fibulin-1 to ADAMTS-1 enzyme-linked immunosorbent binding of a of fibulin-1 to ADAMTS-1, fibulin-1 and ADAMTS-1 was using affinity of As in interacting fibulin-1 to and fibulin-1 fibulin-1 binding was the to the was to the ADAMTS-1 affinity and of of that ADAMTS-1 fibulin-1 and that fibulin-1 performed analyses using and fibulin-1 that ADAMTS-1 fibulin-1 fibulin-1 was found to ADAMTS-1 ADAMTS-1 fibulin-1 not in using these the that fibulin-1 to the biological of is to these proteins the a of and in fibulin-1 and ADAMTS-1 was a in the the of was in the in using and a ADAMTS-1 of fibulin-1 to of the and the of ADAMTS-1 was in the of and is to that was in ADAMTS-1 is not in the that and that ADAMTS-1 is in a of the of the in the is in and of ADAMTS-1 and fibulin-1. was to and and the was in a a fibulin-1 and a to and of a the and ADAMTS-1 of and ADAMTS-1 in the and was performed and of the and in in the the fibulin-1 was a substrate using two fibulin-1 showed cleavage was the of fibulin-1 ADAMTS-1 and the that fibulin-1 fibulin-1 ADAMTS-1 and the using cleavage using fibulin-1 is an that cleavage We fibulin-1 to bind to and the ability of ADAMTS-1 to fibulin-1 As in fibulin-1 was not the that fibulin-1 is not a substrate of is not a substrate and fibulin-1 was ADAMTS-1 and and of using the region of fibulin-1 and to the carboxyl-terminal region of and fibulin-1 was as ADAMTS-1 fibulin-1 and fibulin-1 to of ADAMTS-1, ADAMTS-1 of fibulin-1 was the of fibulin-1 was not ADAMTS-1, that the the function of has been to bind the of aggrecan and two ADAMTS-1 the that fibulin-1 the proteolytic of ADAMTS-1 In the of aggrecan was to the ADAMTS-1 In of proteins and in of the aggrecan the was to enhance ADAMTS-1 was not as as fibulin-1. proteins to enhance ADAMTS-1 proteolytic of the proteins and to an inhibition of ADAMTS-1 cleavage of aggrecan ADAMTS-1 is fibulin-1. of of of aggrecan the proteins in the of of of aggrecan ADAMTS-1 in the of fibulin-1 the in and proteolytic of and of of aggrecan ADAMTS-1 in the of of fibulin-1 of aggrecan of fibulin-1 in the in and the and the the in of aggrecan in the of ADAMTS-1 the cleavage of to a and a the of fibulin-1 the of aggrecan In the of the was is that of ADAMTS-1 the aggrecan to as as in the of fibulin-1. In the aggrecan was and was was the aggrecan cleavage fibulin-1 was of fibulin-1 protein the cleavage was studies showed that the of aggrecan cleavage to the and was to the of fibulin-1 in the of ADAMTS-1 was to as of to ADAMTS-1 Furthermore, the of that of fibulin-1 the of aggrecan and a in the of aggrecan a was of fibulin-1 and not the of proteins aggrecan to In the of not in the aggrecan as a that fibulin-1 the of aggrecan in a that is not to fibulin-1 the of aggrecan, of the proteoglycan ADAMTS-1 a in the of and ADAMTS-1 in the and to is the of aggrecan the ADAMTS-1 in the was the is to ADAMTS-1 of ADAMTS-1 in the of that of the not in of and In ADAMTS-1 of We of aggrecan and ADAMTS-1 in and ADAMTS-1 of was in the as as in the a found fibulin-1 of aggrecan was not in the of ADAMTS-1 using a to aggrecan to ADAMTS-1 aggrecan was in the of ADAMTS-1 in the ADAMTS-1 is a of ADAMTS-1 and of in the of and ADAMTS-1 of to the ADAMTS-1 of the of of to of ADAMTS-1 of aggrecan a and ADAMTS-1 and and aggrecan and in was performed in of of as the of the of the and in in the the in a yeast two-hybrid that fibulin-1 to ADAMTS-1 the in fibulin-1 and and in of these proteins was further confirmed assays, and co-immunoprecipitation, and was in vivo. is that ADAMTS-1 and as the interacting region is to the biological of the of fibulin-1 the of We found that fibulin-1 the cleavage of aggrecan proteoglycan to fibulin-1 affinity the and in these that the of fibulin-1 aggrecan to these proteins in proteolytic two-hybrid been in the of extracellular protein the of matrix in yeast two-hybrid has and the substrate and the is the of of the in matrix has been to In of region has a the of the In a protein in ADAMTS-1 been to interact and to bind to proteins of the of the in and In of the carboxyl-terminal region in the of that the function of the carboxyl-terminal region in was found to bind the carboxyl-terminal binding proteolytic aggrecan to fibulin-1 not the of ADAMTS-1 was found to a substrate the the and fibulin-1 the of found that proteolytic cleavage of aggrecan not to the as fibulin-1 has been the proteolytic of an that binding of the to the substrate in of ADAMTS-1 and fibulin-1 that of and ADAMTS-1 found colocalized in the studies that of fibulin-1 and ADAMTS-1 in the and that ADAMTS-1 not ADAMTS-1 in a fibulin-1 and the of ADAMTS-1 in aggrecan to In a that ADAMTS-1 not in aggrecan in the and a of and In the the as of and is an ADAMTS-1 in the found of aggrecan in the ADAMTS-1 in to to the the ADAMTS-1 is and In as showed of ADAMTS-1 and fibulin-1 fibulin-1 an in the of ADAMTS-1 of the fibulin-1 and of the has been in found that the and fibulin-1 in the of and that has affinity and is a of known to the of matrix the of an the of that is to a in the of the the extracellular matrix extracellular a and metalloprotease extracellular a and metalloprotease of and as and angiogenesis ADAMTS-1 and metalloprotease to a of in the of proteins as and and proteoglycans, aggrecan, and of that ADAMTS-1 and that and In ADAMTS-1 of in the and of in the of ADAMTS-1 and to a of the of ADAMTS-1 biological and of a of that of matrix and of the the of a series of a fibulin-1 to and and to the proteoglycans, aggrecan and fibulin-1 and in the in to in to in a series of found that fibulin-1 to ADAMTS-1 and that fibulin-1 bind to aggrecan, to that fibulin-1 to enhance a was the of ADAMTS-1 was using the and and the and a of proteins was and and and and using a using and and the to in and ADAMTS-1 in a of and to proteins using and to and and of in the using a was protein to of the to fibulin-1 ADAMTS-1 using protein protein and the and proteins using and to and and in the of was and protein was to and protein and the using and ADAMTS-1 and protein and proteins using and to and of ADAMTS-1 and fibulin-1 was and ADAMTS-1 fibulin-1 in binding was binding in ADAMTS-1 fibulin-1 in and and proteins ADAMTS-1 fibulin-1 and performed using substrate and using a ADAMTS-1, in and in and and of fibulin-1 to in and and and in and and and in and and was performed using substrate of was performed using was to of the was in of and using a was and was a of in and a of in a of was of and of proteins a and of and of proteins and to of ADAMTS-1, and in in in and and and in in in in to in to in and to in was using a was in using ADAMTS-1 protein as was and ADAMTS-1 of was ADAMTS-1 and in in a of in using and to the the of was the fibulin-1 was in to fibulin-1 was of ADAMTS-1, ADAMTS-1 in in a of fibulin-1 was in fibulin-1 was of ADAMTS-1 in in a of proteins aggrecan was a is further to the and and of to in the of ADAMTS-1 was in of in of to in and aggrecan was ADAMTS-1 in the of of fibulin-1 as of aggrecan cleavage was using aggrecan ADAMTS-1 was in was and fibulin-1 was and was and of was performed as of was performed and was included a ADAMTS-1 and a the of of ADAMTS-1 of was the region to and a to the of and and was a the to of a and the and a was a was and to the to the and in and identified two and was of ADAMTS-1 was of using a was using that to the and the of was using to the of as and the was performed using of was the of ADAMTS-1 was using the and and the and a of proteins was and and and and using a using and and the to in and ADAMTS-1 in a of and to proteins using and to and and of in the using a was protein to of the to fibulin-1 ADAMTS-1 using protein protein and the and proteins using and to and and in the of was and protein was to and protein and the using and ADAMTS-1 and protein and proteins using and to and of ADAMTS-1 and fibulin-1 was and ADAMTS-1 fibulin-1 in binding was binding in ADAMTS-1 fibulin-1 in and and proteins ADAMTS-1 fibulin-1 and performed using substrate and using a ADAMTS-1, in and in and and of fibulin-1 to in and and and in and and and in and and was performed using substrate of was performed using ADAMTS-1 was to of the was in of and using a was and was a of in and a of in a of was of and of proteins a and of and of proteins and to of ADAMTS-1, and in in in and and and in in in in to in to in and to in was using a was in using ADAMTS-1 protein as was and ADAMTS-1 of was ADAMTS-1 and in in a of in using and to the the of fibulin-1. was the fibulin-1 was in to fibulin-1 was of ADAMTS-1, ADAMTS-1 in in a of fibulin-1 was in fibulin-1 was of ADAMTS-1 in in a of proteins aggrecan was a is further to the and and of to in the of ADAMTS-1 was in of in of to in and aggrecan was ADAMTS-1 in the of of fibulin-1 as of aggrecan cleavage was using aggrecan ADAMTS-1 was in was and fibulin-1 was and was and of was performed as of was performed and was included a ADAMTS-1 and a the of fibulin-1. of ADAMTS-1 of was the region to and a to the of and and was a the to of a and the and a was a was and to the to the and in and identified two and was of ADAMTS-1 was of using a was using that to the and the of was using to the of as and the was performed using of of as an yeast two-hybrid screen of a was to proteins of interacting of the carboxyl-terminal of ADAMTS-1 the two was as a to screen potential binding proteins a that to of a to the of the two and the carboxyl-terminal of the ADAMTS-1 and fibulin-1 was further to fibulin-1 to bind to ADAMTS-1 in not was to ADAMTS-1 of ADAMTS-1 in the of In the was enzyme-linked immunosorbent using As in fibulin-1 to was and ADAMTS-1 was found to bind to fibulin-1 binding affinity these two proteins was was as a has been to interact fibulin-1 ADAMTS-1, and to bind fibulin-1 a of the to a of the binding fibulin-1 to ADAMTS-1 an of as of fibulin-1 binding to As binding was not fibulin-1 and to the that fibulin-1 binding of fibulin-1 to the ADAMTS-1, fibulin-1 binding fibulin-1 and ADAMTS-1 was using affinity of As in interacting fibulin-1 to and fibulin-1 fibulin-1 binding was the to the was to the ADAMTS-1 affinity and of of that ADAMTS-1 fibulin-1 and that fibulin-1 performed analyses using and fibulin-1 that ADAMTS-1 fibulin-1 fibulin-1 was found to ADAMTS-1 ADAMTS-1 fibulin-1 not in using these the that fibulin-1 to the biological of is to these proteins the a of and in fibulin-1 and ADAMTS-1 was a in the the of was in the in using and a ADAMTS-1 of fibulin-1 to of the and the of ADAMTS-1 was in the of and is to that was in ADAMTS-1 is not in the that and that ADAMTS-1 is in a of the of the in the is in and of ADAMTS-1 and fibulin-1. was to and and the was in a a fibulin-1 and a to and of a the and ADAMTS-1 of and ADAMTS-1 in the and was performed and of the and in in the the fibulin-1 was a substrate using two fibulin-1 showed cleavage was the of fibulin-1 ADAMTS-1 and the that fibulin-1 fibulin-1 ADAMTS-1 and the using cleavage using fibulin-1 is an that cleavage We fibulin-1 to bind to and the ability of ADAMTS-1 to fibulin-1 As in fibulin-1 was not the that fibulin-1 is not a substrate of is not a substrate and fibulin-1 was ADAMTS-1 and and of using the region of fibulin-1 and to the carboxyl-terminal region of and fibulin-1 was as ADAMTS-1 fibulin-1 and fibulin-1 to of ADAMTS-1, ADAMTS-1 of fibulin-1 was the of fibulin-1 was not ADAMTS-1, that the the function of has been to bind the of aggrecan and two ADAMTS-1 the that fibulin-1 the proteolytic of ADAMTS-1 In the of aggrecan was to the ADAMTS-1 In of proteins and in of the aggrecan the was to enhance ADAMTS-1 was not as as fibulin-1. proteins to enhance ADAMTS-1 proteolytic of the proteins and to an inhibition of ADAMTS-1 cleavage of aggrecan ADAMTS-1 is fibulin-1. of of of aggrecan the proteins in the of of of aggrecan ADAMTS-1 in the of fibulin-1 the in and proteolytic of and of of aggrecan ADAMTS-1 in the of of fibulin-1 of aggrecan of fibulin-1 in the in and the and the the in of aggrecan in the of ADAMTS-1 the cleavage of to a and a the of fibulin-1 the of aggrecan In the of the was is that of ADAMTS-1 the aggrecan to as as in the of fibulin-1. In the aggrecan was and was was the aggrecan cleavage fibulin-1 was of fibulin-1 protein the cleavage was studies showed that the of aggrecan cleavage to the and was to the of fibulin-1 in the of ADAMTS-1 was to as of to ADAMTS-1 Furthermore, the of that of fibulin-1 the of aggrecan and a in the of aggrecan a was of fibulin-1 and not the of proteins aggrecan to In the of not in the aggrecan as a that fibulin-1 the of aggrecan in a that is not to fibulin-1 the of aggrecan, of the proteoglycan ADAMTS-1 a in the of and ADAMTS-1 in the and to is the of aggrecan the ADAMTS-1 in the was the is to ADAMTS-1 of ADAMTS-1 in the of that of the not in of and In ADAMTS-1 of We of aggrecan and ADAMTS-1 in and ADAMTS-1 of was in the as as in the a found fibulin-1 of aggrecan was not in the of ADAMTS-1 using a to aggrecan to ADAMTS-1 aggrecan was in the of ADAMTS-1 in the ADAMTS-1 is a of ADAMTS-1 and of in the of and ADAMTS-1 of to the ADAMTS-1 of the of of to of ADAMTS-1 of aggrecan a and ADAMTS-1 and and aggrecan and in was performed in of of as the of the of the and in in the the in of as an yeast two-hybrid screen of a was to proteins of interacting of the carboxyl-terminal of ADAMTS-1 the two was as a to screen potential binding proteins a that to of a to the of the two and the carboxyl-terminal of the ADAMTS-1 and fibulin-1 was further to fibulin-1 to bind to ADAMTS-1 in not was to ADAMTS-1 of ADAMTS-1 in the of In the was enzyme-linked immunosorbent using As in fibulin-1 to was and ADAMTS-1 was found to bind to fibulin-1 binding affinity these two proteins was was as a has been to interact fibulin-1 ADAMTS-1, and to bind fibulin-1 a of the to a of the binding fibulin-1 to ADAMTS-1 an of as of fibulin-1 binding to As binding was not fibulin-1 and to the that fibulin-1 binding of fibulin-1 to the ADAMTS-1, fibulin-1 binding fibulin-1 and ADAMTS-1 was using affinity of As in interacting fibulin-1 to and fibulin-1 fibulin-1 binding was the to the was to the performed analyses using and fibulin-1 that ADAMTS-1 fibulin-1 fibulin-1 was found to ADAMTS-1 ADAMTS-1 fibulin-1 not in using these the that fibulin-1 to the biological of is to these proteins the a of and ADAMTS-1 in fibulin-1 and ADAMTS-1 was a in the the of was in the in using and a ADAMTS-1 of fibulin-1 to of the and the of ADAMTS-1 was in the of and is to that was in ADAMTS-1 is not in the that and that ADAMTS-1 is in a of the of the in the is in fibulin-1 was a substrate using two fibulin-1 showed cleavage was the of fibulin-1 ADAMTS-1 and the that fibulin-1 fibulin-1 ADAMTS-1 and the using cleavage using fibulin-1 is an that cleavage We fibulin-1 to bind to and the ability of ADAMTS-1 to fibulin-1 As in fibulin-1 was not the that fibulin-1 is not a substrate of the of fibulin-1 was not ADAMTS-1, that the the function of has been to bind the of aggrecan and two ADAMTS-1 the that fibulin-1 the proteolytic of ADAMTS-1 In the of aggrecan was to the ADAMTS-1 In of proteins and in of the aggrecan the was to enhance ADAMTS-1 was not as as fibulin-1. proteins to enhance ADAMTS-1 proteolytic of the proteins and to an inhibition of ADAMTS-1 the of fibulin-1 the of aggrecan In the of the was is that of ADAMTS-1 the aggrecan to as as in the of fibulin-1. In the aggrecan was and was was the aggrecan cleavage fibulin-1 was of fibulin-1 protein the cleavage was studies showed that the of aggrecan cleavage to the and was to the of fibulin-1 in the of ADAMTS-1 was to as of to ADAMTS-1 Furthermore, the of that of fibulin-1 the of aggrecan and a in the of aggrecan a was of fibulin-1 and not the of proteins aggrecan to In the of not in the aggrecan as a that fibulin-1 the of aggrecan in a that is not to fibulin-1 the of aggrecan, of the proteoglycan ADAMTS-1 ADAMTS-1 a in the of and ADAMTS-1 in the and to is the of aggrecan the ADAMTS-1 in the was the is to ADAMTS-1 of ADAMTS-1 in the of that of the not in of and In ADAMTS-1 of We of aggrecan and ADAMTS-1 in and ADAMTS-1 of was in the as as in the a found fibulin-1 of aggrecan was not in the of ADAMTS-1 using a to aggrecan to ADAMTS-1 aggrecan was in the of ADAMTS-1 in the ADAMTS-1 is a a yeast two-hybrid that fibulin-1 to ADAMTS-1 the in fibulin-1 and and in of these proteins was further confirmed assays, and co-immunoprecipitation, and was in vivo. is that ADAMTS-1 and as the interacting region is to the biological of the of fibulin-1 the of We found that fibulin-1 the cleavage of aggrecan proteoglycan to fibulin-1 affinity the and in these that the of fibulin-1 aggrecan to these proteins in proteolytic two-hybrid been in the of extracellular protein the of matrix in yeast two-hybrid has and the substrate and the is the of of the in matrix has been to In of region has a the of the In a protein in ADAMTS-1 been to interact and to bind to proteins of the of the in and In of the carboxyl-terminal region in the of that the function of the carboxyl-terminal region in was found to bind the carboxyl-terminal binding proteolytic aggrecan to fibulin-1 not the of ADAMTS-1 was found to a substrate the the and fibulin-1 the of found that proteolytic cleavage of aggrecan not to the as fibulin-1 has been the proteolytic of an that binding of the to the substrate in of ADAMTS-1 and fibulin-1 that of and ADAMTS-1 found colocalized in the studies that of fibulin-1 and ADAMTS-1 in the and that ADAMTS-1 not ADAMTS-1 in a fibulin-1 and the of ADAMTS-1 in aggrecan to In a that ADAMTS-1 not in aggrecan in the and a of and In the the as of and is an ADAMTS-1 in the found of aggrecan in the ADAMTS-1 in to to the the ADAMTS-1 is and In as showed of ADAMTS-1 and fibulin-1 fibulin-1 an in the of ADAMTS-1 of the fibulin-1 and of the has been in found that the and fibulin-1 in the of and that has affinity and is a of known to the of matrix the of an the of that is to a in the of the a yeast two-hybrid that fibulin-1 to ADAMTS-1 the in fibulin-1 and and in of these proteins was further confirmed assays, and co-immunoprecipitation, and was in vivo. is that ADAMTS-1 and as the interacting region is to the biological of the of fibulin-1 the of We found that fibulin-1 the cleavage of aggrecan proteoglycan to fibulin-1 affinity the and in these that the of fibulin-1 aggrecan to these proteins in proteolytic two-hybrid been in the of extracellular protein the of matrix in yeast two-hybrid has and the substrate and the is the of of the in matrix has been to In of region has a the of the In a protein in ADAMTS-1 been to interact and to bind to proteins of the of the in and In of the carboxyl-terminal region in the of that the function of the carboxyl-terminal region in was found to bind the carboxyl-terminal binding proteolytic aggrecan to fibulin-1 not the of ADAMTS-1 was found to a substrate the the and fibulin-1 the of found that proteolytic cleavage of aggrecan not to the as fibulin-1 has been the proteolytic of an that binding of the to the substrate in of ADAMTS-1 and fibulin-1 that of and ADAMTS-1 found colocalized in the studies that of fibulin-1 and ADAMTS-1 in the and that ADAMTS-1 not ADAMTS-1 in a fibulin-1 and the of ADAMTS-1 in aggrecan to In a that ADAMTS-1 not in aggrecan in the and a of and In the the as of and is an ADAMTS-1 in the found of aggrecan in the ADAMTS-1 in to to the the ADAMTS-1 ADAMTS-1 is and In as showed of ADAMTS-1 and fibulin-1 fibulin-1 an in the of ADAMTS-1 of the fibulin-1 and of the has been in found that the and fibulin-1 in the of and that has affinity and is a of known to the of matrix the of an the of that is to a in the of the We aggrecan