Key points are not available for this paper at this time.
Lysyl oxidase catalyzes the final enzymatic step required for collagen and elastin cross-linking in extracellular matrix biosynthesis. Pro-lysyl oxidase is processed by procollagen C-proteinase activity, which also removes the C-propeptides of procollagens I–III. The Bmp1 gene encodes two procollagen C-proteinases: bone morphogenetic protein 1 (BMP-1) and mammalian Tolloid (mTLD). Mammalian Tolloid-like (mTLL)-1 and -2 are two genetically distinct BMP-1-related proteinases, and mTLL-1 has been shown to have procollagen C-proteinase activity. The present study is the first to directly compare pro-lysyl oxidase processing by these four related proteinases. In vitro assays with purified recombinant enzymes show that all four proteinases productively cleave pro-lysyl oxidase at the correct physiological site but that BMP-1 is 3-, 15-, and 20-fold more efficient than mTLL-1, mTLL-2, and mTLD, respectively. To more directly assess the roles of BMP-1 and mTLL-1 in lysyl oxidase activation by connective tissue cells, fibroblasts cultured from Bmp1-null,Tll1-null, and Bmp1/Tll1 double null mouse embryos, thus lacking BMP-1/mTLD, mTLL-1, or all three enzymes, respectively, were assayed for lysyl oxidase enzyme activity and for accumulation of pro-lysyl oxidase and mature ∼30-kDa lysyl oxidase. Wild type cells or cells singly null for Bmp1 orTll1 all produced both pro-lysyl oxidase and processed lysyl oxidase at similar levels, indicating apparently normal levels of processing, consistent with enzyme activity data. In contrast, double null Bmp1/Tll1 cells produced predominantly unprocessed 50-kDa pro-lysyl oxidase and had lysyl oxidase enzyme activity diminished by 70% compared with wild type, Bmp1-null, andTll1-null cells. Thus, the combination of BMP-1/mTLD and mTLL-1 is shown to be responsible for the majority of processing leading to activation of lysyl oxidase by murine embryonic fibroblasts, whereas in vitro studies identify pro-lysyl oxidase as the first known substrate for mTLL-2. Lysyl oxidase catalyzes the final enzymatic step required for collagen and elastin cross-linking in extracellular matrix biosynthesis. Pro-lysyl oxidase is processed by procollagen C-proteinase activity, which also removes the C-propeptides of procollagens I–III. The Bmp1 gene encodes two procollagen C-proteinases: bone morphogenetic protein 1 (BMP-1) and mammalian Tolloid (mTLD). Mammalian Tolloid-like (mTLL)-1 and -2 are two genetically distinct BMP-1-related proteinases, and mTLL-1 has been shown to have procollagen C-proteinase activity. The present study is the first to directly compare pro-lysyl oxidase processing by these four related proteinases. In vitro assays with purified recombinant enzymes show that all four proteinases productively cleave pro-lysyl oxidase at the correct physiological site but that BMP-1 is 3-, 15-, and 20-fold more efficient than mTLL-1, mTLL-2, and mTLD, respectively. To more directly assess the roles of BMP-1 and mTLL-1 in lysyl oxidase activation by connective tissue cells, fibroblasts cultured from Bmp1-null,Tll1-null, and Bmp1/Tll1 double null mouse embryos, thus lacking BMP-1/mTLD, mTLL-1, or all three enzymes, respectively, were assayed for lysyl oxidase enzyme activity and for accumulation of pro-lysyl oxidase and mature ∼30-kDa lysyl oxidase. Wild type cells or cells singly null for Bmp1 orTll1 all produced both pro-lysyl oxidase and processed lysyl oxidase at similar levels, indicating apparently normal levels of processing, consistent with enzyme activity data. In contrast, double null Bmp1/Tll1 cells produced predominantly unprocessed 50-kDa pro-lysyl oxidase and had lysyl oxidase enzyme activity diminished by 70% compared with wild type, Bmp1-null, andTll1-null cells. Thus, the combination of BMP-1/mTLD and mTLL-1 is shown to be responsible for the majority of processing leading to activation of lysyl oxidase by murine embryonic fibroblasts, whereas in vitro studies identify pro-lysyl oxidase as the first known substrate for mTLL-2. procollagen C-proteinase bone morphogenetic protein mammalian Tolloid mammalian Tolloid-like polyacrylamide gel electrophoresis Lysyl oxidase catalyzes the oxidative deamination of peptidyl-lysine in elastin precursors and lysine and hydroxylysine residues in collagen to form peptidyl-α-aminoadipic-δ-semialdehyde and peptidyl-δ-hydroxy-α-aminoadipic-δ-semialdehyde residues, respectively. These aldehydes then undergo nonenzymatic reactions resulting in the cross-linkages known to be critical in the formation of mature functional insoluble elastin and collagens (1Kagan H. Mecham R.P. Biology and Regulation of Extracellular Matrix: Regulation of Matrix Accumulation. 1. Academic Press, Orlando1986: 321-398Google Scholar, 2Kagan H.M. Trackman P.C. Am. J. Respir. Cell Mol. Biol. 1991; 5: 206-210Crossref PubMed Scopus (281) Google Scholar). Lysyl oxidase is a copper-dependent enzyme and is specifically inhibited by β-aminopropionitrile. Connective tissue abnormalities known as osteolathyrism result fromin vivo inhibition of lysyl oxidase activity by β-aminopropionitrile feeding (3Siegel R.C. Int. Rev. Connect. Tis. Res. 1979; 8: 73-118Crossref PubMed Google Scholar). The abnormalities include malformed and weak bones, as well as increased development of hernias (4Selye H. Rev. Can. Biol. 1957; 16: 3-82Google Scholar). Lysyl oxidase is synthesized as a 50-kDa glycoprotein and is processed extracellularly to produce the ∼30-kDa molecular form known to be active (5Trackman P.C. Bedell-Hogan D. Tang J. Kagan H.M. J. Biol. Chem. 1992; 267: 8666-8671Abstract Full Text PDF PubMed Google Scholar). The sequence of the proteolytic processing site in pro-lysyl oxidase resembles that of the fibrillar procollagen C-terminal pro-peptide processing sites cleaved by procollagen C-proteinase (PCP)1 (6Cronshaw A.D. Fothergill-Gilmore L.A. Hulmes D.J.S. Biochem. J. 1995; 306: 279-284Crossref PubMed Scopus (77) Google Scholar). Moreover, preparations highly enriched in PCP activity have been shown to process pro-lysyl oxidase at the correct physiological site (7Panchenko M.V. Stetler-Stevenson W.G. Trubetskoy O.V. Gacheru S.N. Kagan H.M. J. Biol. Chem. 1996; 271: 7113-7119Abstract Full Text Full Text PDF PubMed Scopus (172) Google Scholar). In mammals, PCP activity is provided by products of the Bmp1gene (8Kessler E. Takahara K. Biniaminov L. Brusel M. Greenspan D.S. Science. 1996; 271: 360-362Crossref PubMed Scopus (458) Google Scholar, 9Li S.W. Sieron A.L. Fertala A. Hojima Y. Arnold W.V. Prockop D.J. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: 5127-5130Crossref PubMed Scopus (202) Google Scholar, 10Scott I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar), which encodes alternatively spliced mRNAs for the proteins bone morphogenetic protein 1 (BMP-1) and mammalian Tolloid (mTLD) (11Takahara K. Kessler E. Biniaminov L. Brusel M. Eddy R.L. Jani-Sait S. Shows T.B. Greenspan D.S. J. Biol. Chem. 1994; 269: 26280-26285Abstract Full Text PDF PubMed Google Scholar). The mTLD protein product contains a longer C terminus than BMP-1, resulting in a domain to that of protein The in BMP-1, mTLD, Tolloid include the domain and and that to extracellular proteins D.J. Sieron A.L. S.W. Matrix Biol. 16: PubMed Scopus Google Scholar, D.J. Kessler E. Matrix Biol. 16: PubMed Scopus Google Tolloid in formation by the protein which with the protein M. K. Cho K.W. Full Text Full Text PDF PubMed Scopus Google Scholar). BMP-1 a similar in mammalian the which and the I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar, Full Text Full Text PDF PubMed Scopus Google Scholar). mammalian BMP-1 and mTLD both are BMP-1 indicating that Bmp1 gene products have substrate and I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). The mammalian Bmp1 gene is a of a and two genetically distinct mammalian proteinases, mammalian Tolloid-like 1 and and have been I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). highly similar and domain the substrate of BMP-1, mTLD, mTLL-1, and BMP-1, mTLD, and mTLL-1 all have PCP activity. contrast, to process fibrillar BMP-1 and mTLL-1 cleave whereas mTLD and cleave I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). BMP-1 and mTLL-1 process whereas I.C. Imamura Y. Pappano W.N. A.D. Greenspan D.S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). has to be in with as pro-lysyl oxidase. lysyl oxidase is critical for of mature and functional extracellular is to a of the BMP-1-related proteinases in processing pro-lysyl oxidase in connective and cells or functional for activity BMP-1-related null for the Bmp1 gene have of and of the and but have normal and Y. L. Takahara K. Greenspan D.S. 1996; PubMed Google Scholar). In fibroblasts from insoluble and collagen are normal in processing is resulting in accumulation of that the and in of processed collagen Y. L. Takahara K. Greenspan D.S. 1996; PubMed Google Scholar). These that in and fibroblasts, related proteinases are to but for the of the PCP activity provided by BMP-1 and the that and abnormalities in similar to (4Selye H. Rev. Can. Biol. 1957; 16: 3-82Google Scholar). is by a in lysyl oxidase enzyme activity (1Kagan H. Mecham R.P. Biology and Regulation of Extracellular Matrix: Regulation of Matrix Accumulation. 1. Academic Press, Orlando1986: 321-398Google Scholar, R.C. Int. Rev. Connect. Tis. Res. 1979; 8: 73-118Crossref PubMed Google Scholar). These the that lysyl oxidase enzyme activity be in the of a functional have been and the null is embryonic T.G. I.C. J. Greenspan D.S. 1999; PubMed Google Scholar). In embryos, the embryonic has abnormalities predominantly in and These in with in the of the in T.G. I.C. J. Greenspan D.S. 1999; PubMed Google Scholar), the of more in and functional the products of the two The lysyl oxidase processing of the BMP-1-related proteinases have been has the enzyme responsible for processing of pro-lysyl vivo been The present study pro-lysyl oxidase processing in vitro by all four known mammalian BMP-1 mTLD, mTLL-1, and In lysyl oxidase enzyme activity and processing are in mouse fibroblasts null for for the mTLL-1 or for both from in show that BMP-1, mTLD, mTLL-1, and all process pro-lysyl oxidase at the correct physiological site but that the BMP-1 and mTLL-1 enzymes are the Moreover, from in and from the that the Bmp1 and the majority of activity for the processing of pro-lysyl oxidase and the of lysyl oxidase enzyme activity. The studies also identify pro-lysyl oxidase as the first known substrate for mTLL-2. BMP-1, mTLD, mTLL-1, and were produced as C-terminal proteins in cells and purified by the as in 10Scott I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google enzyme a with and of enzyme to be or The of enzyme by of enzymes and known of proteins to and I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). The of enzymes known type procollagen and were to with pro-lysyl oxidase I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). The oxidase protein produced and purified as Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). lysyl oxidase were purified mature lysyl oxidase as P.C. A. Tang Kagan H.M. PubMed Scopus Google and were provided by M. Kagan of The were purified lysyl oxidase protein Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). of oxidase recombinant lysyl oxidase protein to and the were purified to the The of the purified lysyl oxidase by the lysyl oxidase and were is sequence lysyl oxidase and the the purified oxidase both mature lysyl oxidase and the protein Thus, by lysyl oxidase and the protein to for of both the lysyl oxidase and the protein respectively, of the recombinant protein substrate by oxidase protein with of recombinant BMP-1, mTLD, mTLL-1, or for or at in a final of of The reactions all and to proteinases. The reactions were by of and for were to and H. J. Proc. Natl. Acad. Sci. U. S. A. 1979; PubMed Scopus Google to and the were with in The were with purified oxidase at a of at The were and then with to the The were The of protein substrate and lysyl oxidase product in by in a with and the of the were The pro-lysyl oxidase then from reactions in which of protein to product or studies have shown that pro-lysyl oxidase processing activity assays as are as a of of in reactions in which to of protein substrate to product is Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). sequence of reactions of as were at for The reactions but all The reactions were by to to and with protein were and to by by and at the from Bmp1 Bmp1 double null and were as Y. L. Takahara K. Greenspan D.S. 1996; PubMed Google Scholar, T.G. I.C. J. Greenspan D.S. 1999; PubMed Google Scholar). The cells were more than and were in tissue in and Lysyl oxidase enzyme activity in a recombinant substrate D. Trackman J. Kagan H. J. Biol. Chem. Full Text PDF PubMed Google Scholar). were assayed in in a final of 1 and of in the and of β-aminopropionitrile. The reactions were for at by The of in in of by of enzyme activity are as β-aminopropionitrile of to enzyme activity were from Biochem. PubMed Scopus Google Scholar). tissue of wild type, Bmp1 and double null fibroblasts were to The cells were then in and for The were then with and cultured for proteins were by with and in levels of proteins from were with lysyl oxidase and with (5Trackman P.C. Bedell-Hogan D. Tang J. Kagan H.M. J. Biol. Chem. 1992; 267: 8666-8671Abstract Full Text PDF PubMed Google Scholar, Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar), and then to and oxidase proteins were and as substrate in assays to assess pro-lysyl oxidase proteolytic processing activity (7Panchenko M.V. Stetler-Stevenson W.G. Trubetskoy O.V. Gacheru S.N. Kagan H.M. J. Biol. Chem. 1996; 271: 7113-7119Abstract Full Text Full Text PDF PubMed Scopus (172) Google Scholar, Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). shown in the of the protein of the and the C-terminal is the lysyl oxidase proteolytic processing by two products are mature lysyl oxidase and a oxidase of products lysyl oxidase and of lysyl oxidase processing activity Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). protein processing by purified recombinant BMP-1, mTLD, mTLL-1, and of purified enzyme with of protein for at and products were to and with lysyl oxidase as protein and a and with the lysyl oxidase Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). product in the of a recombinant enzyme in of a product by the lysyl oxidase these BMP-1 and mTLL-1 to of protein and of of whereas mTLD and in of protein and of of of the of and in by BMP-1 and mTLL-1 and by mTLD and mTLL-2, respectively. were in a of four with two preparations of of lysyl oxidase protein is with to both the of and the of activity the of is or the in the present study and as Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). Thus, were for with the of activity for all four for in for all enzymes, as BMP-1 of the oxidase and mTLL-1 activity then for and the were as of protein of enzyme The in are from shown in and and and The show that BMP-1 the lysyl oxidase protein with the compared with the from four in which were for or all of protein to lysyl oxidase by BMP-1 compared with the enzymes, with similar to the shown in A. The in are from assays in the of the and by these that BMP-1 is the efficient pro-lysyl oxidase processing These are of all of the oxidase processing activity by recombinant BMP-1, mTLD, mTLL-1, and oxidase processing of oxidase protein with of recombinant enzyme for or at and were to and with lysyl oxidase The of of protein to lysyl oxidase by of both in Pro-lysyl oxidase processing enzyme then from reactions in which or Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). are from of shown in in a of oxidase protein with of recombinant enzyme for or at and were to and with lysyl oxidase The of of protein to lysyl oxidase by of both in Pro-lysyl oxidase processing enzyme then from reactions in which or Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). are from of shown in sequence were the products from lysyl oxidase protein and BMP-1, mTLD, mTLL-1, and mTLL-2, respectively. The terminus of to the known terminus of the tissue form of mature active lysyl oxidase with the of from all four product from reactions Thus, the of processing of the lysyl oxidase protein by the four enzymes the product in all to the product of pro-lysyl oxidase. The that the two products of the Bmp1 and mTLD, and products from the genetically distinct and mTLL-1 and mTLL-2, respectively, all process pro-lysyl oxidase in of the to that the be to diminished lysyl oxidase activity, at in as and The the pro-lysyl oxidase processing activity of BMP-1, that lysyl oxidase processing and vivo to a products of gene and to BMP-1 in To levels of lysyl oxidase enzyme activity were first from of fibroblasts from wild type, Bmp1 and double null The that lysyl oxidase enzyme activity by the double null embryonic fibroblasts is of that in cells from wild type, and cells, Thus, cells null for both the Bmp1 produced a in levels of lysyl oxidase enzyme activity. of molecular of lysyl oxidase then by in wild type, and double null cells to 50-kDa pro-lysyl oxidase processing diminished in the double null cells. The cells were cultured to and then cultured in for proteins were then with oxidase or and to and shown in unprocessed 50-kDa lysyl oxidase at levels in from double null cells compared with the of the 50-kDa lysyl oxidase that the double null cells 3-, and more 50-kDa lysyl oxidase than wild type, Bmp1 cells, respectively. The pro-lysyl oxidase protein from from all is predominantly the 50-kDa form of protein but also the consistent with studies in cells and murine cells (5Trackman P.C. Bedell-Hogan D. Tang J. Kagan H.M. J. Biol. Chem. 1992; 267: 8666-8671Abstract Full Text PDF PubMed Google Scholar, Trackman P.C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). similar levels of ∼30-kDa lysyl oxidase were in all four but in double null cells the of is and more than in the three that the in the double null processed lysyl oxidase or a of and lysyl oxidase. is consistent with lysyl oxidase enzyme activity that diminished lysyl oxidase activity in Bmp1/Tll1 double null oxidase enzyme activity in of wild type, and Bmp1 and double null murine embryonic oxidase wild oxidase enzyme activity in from to from the are the of from three of 1 and preparations of recombinant The are of of in lysyl oxidase activity and wild type and wild type or Bmp1 and wild type In double null lysyl oxidase enzyme activity than wild type whereas activity in Bmp1 than wild type in a Lysyl oxidase enzyme activity in from to from the are the of from three of 1 and preparations of recombinant The are of of in lysyl oxidase activity and wild type and wild type or Bmp1 and wild type In double null lysyl oxidase enzyme activity than wild type whereas activity in Bmp1 than wild type the the that and produce efficient pro-lysyl oxidase processing enzymes, and that these enzymes to pro-lysyl oxidase processing and development of lysyl oxidase enzyme activity in murine embryonic These a for mTLL-1 in pro-lysyl oxidase processing and In that also to pro-lysyl oxidase to the that proteinases with PCP activity be in and a of extracellular critical for extracellular matrix The two are that enzymes with PCP activity have extracellular and that the Bmp1 which is related to that morphogenetic in encodes BMP-1 and mTLD, two products with PCP activity. In to the processing of procollagen and preparations highly enriched in PCP activity process pro-lysyl oxidase (7Panchenko M.V. Stetler-Stevenson W.G. Trubetskoy O.V. Gacheru S.N. Kagan H.M. J. Biol. Chem. 1996; 271: 7113-7119Abstract Full Text Full Text PDF PubMed Scopus (172) Google Scholar). BMP-1 has been shown to to from I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar, Full Text Full Text PDF PubMed Scopus Google and process collagen Y. Steiglitz B.M. Greenspan D.S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), S. I.C. Takahara K. M. S. Greenspan D.S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), and I.C. Imamura Y. Pappano W.N. A.D. Greenspan D.S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The of a activity that a of leading to a is well known in in which extracellular matrix is by or or H. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar). The of or extracellular enzymatic of enzymes as with PCP activity. with proteinases as the a of to for proteinases, enzyme be by in a and The present study the of proteinases by the Bmp1gene and of two highly similar proteinases mammalian to process pro-lysyl In the of the of proteinases by the Bmp1 and respectively, lysyl oxidase activity and in fibroblasts cultured from wild type, Bmp1-null,Tll1-null, and Bmp1/Tll1 double null mouse both in with recombinant proteins and cultured cells from genetically assays to assess the roles of the procollagen in lysyl oxidase In vitro were with recombinant enzymes, all of which were in the mammalian all of which the C-terminal and all of which were purified by the I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). Moreover, the oxidase protein a lysyl oxidase Y. L. Takahara K. Greenspan D.S. 1996; PubMed Google Scholar), the proteolytic processing site is and pro-lysyl oxidase (6Cronshaw A.D. Fothergill-Gilmore L.A. Hulmes D.J.S. Biochem. J. 1995; 306: 279-284Crossref PubMed Scopus (77) Google Scholar). Thus, the of assays of processing of the lysyl oxidase substrate by BMP-1-related proteinases, of which in the a to pro-lysyl oxidase processing activity. The of the is by the the in vitro and from studies of mouse fibroblasts, all of which the that BMP-1 and mTLL-1 the majority of processing leading to the activation of lysyl oxidase. The in vitro that BMP-1, mTLD, mTLL-1, and enzymes all process pro-lysyl oxidase at the physiological site but in pro-lysyl oxidase processing is that BMP-1, which has the PCP activity, is also the efficient pro-lysyl oxidase processing The of BMP-1 processing of both procollagens and pro-lysyl oxidase BMP-1 as a activity in the of functional insoluble of BMP-1, mTLD, mTLL-1, and in BMP-1 and mTLD to be at levels whereas mTLL-1 more to or and in I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). The Bmp1 gene is also at levels in M. Dev. Biol. 1994; PubMed Scopus Google Scholar). Thus, of of BMP-1 and mTLD in highly are also consistent with a in lysyl oxidase and mature collagen biosynthesis. The more of mTLL-1 and roles for these products in the normal development of extracellular in is that is to process lysyl oxidase but to process procollagens or at all I.C. Imamura Y. Pappano W.N. A.D. Greenspan D.S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). Thus, of the procollagen C-proteinase lysyl oxidase is in that is processed by mTLL-2. that lysyl cross-linking is critical for the of mature connective of for lysyl oxidase activation two proteins are from the Bmp1 gene by BMP-1 and mTLD K. Greenspan D.S. J. Biol. Chem. 1994; 269: Full Text PDF PubMed Google Scholar). is of that mTLD procollagens and pro-lysyl oxidase than is also that mTLD cleave whereas BMP-1 I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). The mTLD protein is longer than the BMP-1 protein and contains two and In to activity, these sites for extracellular matrix for to mTLD E. of Academic Press, Scholar). is that mTLD and BMP-1 proteins are in and mouse K. Greenspan D.S. J. Biol. Chem. 1994; 269: Full Text PDF PubMed Google Scholar). The in vitro that the BMP-1 is a highly efficient pro-lysyl oxidase processing enzyme to the that BMP-1 be the lysyl oxidase processing activity in both cultured cells and in The connective tissue in Bmp1 and development of bones, be in by normal lysyl oxidase activity, in at in these Y. L. Takahara K. Greenspan D.S. 1996; PubMed Google Scholar). The from studies in cultured cells were in that cells processed lysyl oxidase and produced similar lysyl oxidase enzyme activity compared with wild type cells. lysyl oxidase enzyme activity and wild type levels of 50-kDa pro-lysyl oxidase and of ∼30-kDa mature lysyl oxidase in Bmp1 cells. These that enzymes as mTLL-1 for the of BMP-1 mTLD in these that mTLL-1 and mRNAs are in mouse fibroblasts I.C. Imamura Y. Pappano W.N. A.D. Greenspan D.S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). double null cells produce levels of lysyl oxidase activity diminished by 70% and highly levels of the 50-kDa lysyl oxidase the that both BMP-1 and mTLL-1 enzymes are in processing pro-lysyl oxidase and that mTLL-1 to pro-lysyl oxidase processing in the of studies the and of in processing lysyl oxidase be in or a lysyl oxidase processing enzyme for the lysyl oxidase processing and lysyl oxidase enzyme activity in the Bmp1/Tll1 double null murine embryonic with a of and of the of the roles of these gene products in to the lysyl oxidase activity in Bmp1/Tll1 double null cells be that lysyl oxidase enzyme activity be produced by at In of a study has shown that murine that produce or lysyl oxidase protein produce lysyl oxidase activity H. Kessler E. Trackman P.C. J. Res. PubMed Scopus Google Scholar). Moreover, lysyl have been and to and K. S. J. Biol. Chem. Full Text PDF PubMed Google Scholar, H. L. K. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar, K. PubMed Scopus Google Scholar, A. Res. 1999; Google Scholar, H. J. H. S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), whereas the lysyl oxidase gene to Trackman P.C. Kagan H.M. Eddy R.L. Shows T.B. 1992; PubMed Scopus Google Scholar). The of all lysyl oxidase that proteolytic the lysyl oxidase sequence contains a sequence similar to the and present in and procollagen (6Cronshaw A.D. Fothergill-Gilmore L.A. Hulmes D.J.S. Biochem. J. 1995; 306: 279-284Crossref PubMed Scopus (77) Google Scholar). These the that lysyl oxidase activity from or more lysyl of BMP-1-related enzymes and for the lysyl oxidase enzyme activity in cells cultured from be that the in and from the more highly procollagen and pro-lysyl oxidase I.C. Blitz I.L. Pappano W.N. Imamura Y. Clark T.G. Steiglitz B.M. Thomas C.L. Maas S.A. Takahara K. Cho K.W. Greenspan D.S. Dev. Biol. 1999; 213: 283-300Crossref PubMed Scopus (233) Google Scholar). The site in collagen also has and in the Y. Steiglitz B.M. Greenspan D.S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). Thus, the pro-lysyl precursors also processing by BMP-1 BMP-1-related proteinases at a the and of lysyl oxidase and the lysyl proteins are to of functional studies of pro-lysyl processing enzymes distinct from the BMP-1 In a of the substrate of BMP-1 and of related enzymes to the roles of of proteinases. and for Bmp1
Üzel et al. (Fri,) studied this question.
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