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• W2000783985 abstract "Nucleus pulposus, the central zone of the intervertebral disc, is gel-like and has a similar collagen phenotype to that of hyaline cartilage. Amino-terminal protein sequence analysis of the α1(IX)COL3 domain purified from bovine nucleus pulposus gave a different sequence to that of the long α1(IX) transcript expressed in hyaline cartilage and matched the predicted sequence of short α1(IX). The findings indicate that the matrix of bovine nucleus pulposus contains only the short form of α1(IX) that lacks the NC4 domain. The sequence encoded by exon 7, predicted from human COL9A1, is absent from both short and long forms of α1(IX) from bovine nucleus pulposus and articular cartilage. A structural analysis of the cross-linking sites occupied in type IX collagen from nucleus pulposus showed that usage of the short α1(IX) transcript in disc tissue had no apparent effect on cross-linking behavior. As in cartilage, type IX collagen of nucleus pulposus was heavily cross-linked to type II collagen and to other molecules of type IX collagen with a similar site occupancy. Nucleus pulposus, the central zone of the intervertebral disc, is gel-like and has a similar collagen phenotype to that of hyaline cartilage. Amino-terminal protein sequence analysis of the α1(IX)COL3 domain purified from bovine nucleus pulposus gave a different sequence to that of the long α1(IX) transcript expressed in hyaline cartilage and matched the predicted sequence of short α1(IX). The findings indicate that the matrix of bovine nucleus pulposus contains only the short form of α1(IX) that lacks the NC4 domain. The sequence encoded by exon 7, predicted from human COL9A1, is absent from both short and long forms of α1(IX) from bovine nucleus pulposus and articular cartilage. A structural analysis of the cross-linking sites occupied in type IX collagen from nucleus pulposus showed that usage of the short α1(IX) transcript in disc tissue had no apparent effect on cross-linking behavior. As in cartilage, type IX collagen of nucleus pulposus was heavily cross-linked to type II collagen and to other molecules of type IX collagen with a similar site occupancy. The collagen phenotype of hyaline cartilage is complex; at least seven distinct collagen types have been identified, of which types II, XI, and IX are the cartilage-specific molecules. In addition, significant amounts of collagen types III, VI, XII, and XIV have been identified in cartilage matrix (1Eyre D. Arthritis Res. 2002; 4: 30-35Google Scholar, 2Ayad S. Marriott A. Morgan K. Grant M.E. Biochem. J. 1989; 262: 753-761Google Scholar, 3Watt S.L. Lunstrum G.P. McDonough A.M. Keene D.R. Burgeson R.E. Morris N.P. J. Biol. Chem. 1992; 267: 20093-20099Google Scholar). Type IX collagen is a structural matrix component that is most abundant in hyaline cartilages of the developing skeleton. It functions as an adhesion protein in the extracellular matrix where it becomes covalently cross-linked to the surface of type II collagen fibrils and most concentrated on thin fibrils of the pericellular domain (4Mendler M. Eich-Bender S.G. Vaughan L. Winterhalter K.H. Bruckner P. J. Cell Biol. 1989; 108: 191-197Google Scholar, 5Wu J.-J. Woods P.E. Eyre D.R. J. Biol. Chem. 1992; 267: 23007-23014Google Scholar, 6Vaughan L. Mendler M. Huber S. Bruckner P. Winterhalter K.H. Irwin M.I. Mayne R. J. Cell Biol. 1988; 106: 991-997Google Scholar). The molecule is a heterotrimer of genetically distinct α1(IX), α2(IX), and α3(IX) chains, which form three triple-helical segments, COL1, 1The abbreviations used are: COL, collagenous; NC, non-collagenous; HPLC, high performance liquid chromatography; PVDF, polyvinylidene difluoride; N- and C-telopeptides, short sequences that lack the (Gly-XY)n repeat and from the amino and carboxyl ends of various collagen chains. COL2, and COL3 and four non-helical domains, NC1–NC4. The largest, NC4 on α1(IX), is basically charged, suggesting that it may serve to interact with proteoglycans (7Vasios G. Nishimura I. Konomi H. van der Rest M. Ninomiya Y. Olsen B.R. J. Biol. Chem. 1988; 263: 2324-2329Google Scholar). Recent reports have linked genetic polymorphisms to risk of disc degeneration (8Annunen S. Paassilta P. Lohiniva J. Perala M. Pihlajamaa T. Karppinen J. Tervonen O. Kroger H. Lahde S. Vanharanta H. Ryhanen L. Goring H.H.H. Ott J. Prockop D.J. Ala-Kokko L. Science. 1999; 285: 409-412Google Scholar, 9Paassilta P. Lohiniva J. Goring H.H.H. Perala M. Raina S.S. Karppinen J. Hakala M. Palm T. Kroger H. Kaitila I. Vanharanta H. Ott J. Ala-Kokko L. J. Am. Med. Assoc. 2001; 285: 1843-1849Google Scholar). Specifically, tryptophan substitutions at position 326 of the α2(IX) chain, and position 103 of the α3(IX) chain, were associated with increased risk of human lumbar disc degeneration and chronic sciatica but not knee osteoarthritis (8Annunen S. Paassilta P. Lohiniva J. Perala M. Pihlajamaa T. Karppinen J. Tervonen O. Kroger H. Lahde S. Vanharanta H. Ryhanen L. Goring H.H.H. Ott J. Prockop D.J. Ala-Kokko L. Science. 1999; 285: 409-412Google Scholar, 9Paassilta P. Lohiniva J. Goring H.H.H. Perala M. Raina S.S. Karppinen J. Hakala M. Palm T. Kroger H. Kaitila I. Vanharanta H. Ott J. Ala-Kokko L. J. Am. Med. Assoc. 2001; 285: 1843-1849Google Scholar). Nucleus pulposus, the gel-like central zone of the young intervertebral disc, has a similar collagen phenotype to that of hyaline cartilage, with types II, IX, and XI collagens being the principal fibrillar components (10Eyre D.R. Matsui Y. Wu J.-J. Biochem. Soc. Trans. 2002; 30: 844-848Google Scholar). The association of the tryptophan polymorphisms with clinical conditions involving discs but not synovial joint cartilages implies that the molecular character of collagen IX in intervertebral disc collagen may differ from that in hyaline cartilage. The properties of collagen IX from disc tissue have not been fully characterized. The human α1(IX) gene gives rise to two mRNA transcripts, a long and a short form (11Muragaki Y. Kimura T. Ninomiya Y. Olsen B.R. Eur. J. Biochem. 1990; 192: 703-708Google Scholar). The long form is expressed in hyaline cartilages and the product has a globular NH2-terminal domain, NC4. Short α1(IX), expressed in embryonic chick cornea (12Nishimura I. Muragaki Y. Olsen B.R. J. Biol. Chem. 1989; 264: 20033-20041Google Scholar), is transcribed from an alternative start site and lacks the NC4 domain. A second promoter and an alternative exon 1, located in the intron between exons 6 and 7 of COL9A1, are used to express the short form of α1(IX). In the present study, by NH2-terminal sequence analysis of COL3(IX) domains isolated from the matrix of bovine nucleus pulposes and articular cartilage, we show that nucleus pulposus contains exclusively the short form of α1(IX). The covalent cross-linking properties of nucleus pulposus type IX collagen were also characterized to determine any consequences of having short instead of long α1(IX). Preparation of Type IX Collagen—Articular cartilage was dissected from knee joints and nucleus pulposus from lumbar spines of 3-month-old calves. Tissue slices were extracted in 4 m guanidine HCl, 0.05 m Tris-HCl, pH 7.4, at 4 °C for 24 h to remove proteoglycans and other matrix proteins then washed thoroughly with water and freeze-dried. Cross-linked collagens were solubilized by digesting the washed residues with pepsin at 4 °C. Pepsin digests were fractionated into collagen types II, XI, and IX (COL1 and COL2 trimers) by precipitation at 0.7, 1.2, and 2.0 m NaCl, respectively (13Wu J.-J. Eyre D.R. Biochem. Biophys. Res. Commun. 1984; 123: 1033-1039Google Scholar). The COL3 domains of type IX collagen were then isolated from the 4.0 m NaCl precipitated fraction. Column Chromatography—Pepsin-solubilized type IX collagen COL1 and COL2 trimers were resolved on a C8 reverse-phase column (Brownlee Aquapore RP-300; 4.6 mm × 25 cm) with a linear gradient (23–38%) of solvent B in A over 45 min at a flow rate of 1 ml/min collecting 1-ml fractions. Solvent A was 0.1% trifluoroacetic acid (v/v) in water, and solvent B was 0.085% trifluoroacetic acid (v/v) in acetonitrile:n-propyl alcohol (3:1, v/v). Eluent was monitored for 220 nm absorbance, and aliquots of collected fractions (100 μl each) were dried and analyzed by SDS-PAGE. The disulfide-bounded COL2 trimer was reduced, and cysteines were carboxymethylated with iodoacetate before rechromatographing the individual chains on the same column with a linear gradient (15–40%) of solvent B in A over 50 min. The COL3 domains of type IX collagen were isolated from the 4.0 m NaCl precipitated fraction by molecular sieve chromatography on a Bio-Gel A5m column (1.5 × 170 cm, 200–400 mesh, Bio-Rad) eluted with a 0.05 m Tris-HCl buffer, pH 7.5, containing 2 m guanidine HCl, at a flow rate of 4 ml/h, collecting 2.0-ml fractions. Aliquots of collected fractions were desalted and analyzed by SDS-PAGE. Fractions containing the type IX collagen COL3 domain were pooled. The individual COL3(IX) chains were then resolved by HPLC on a C8 reverse-phase column with a linear gradient (15–40%) of solvent B in A over 50 min followed by SDS-PAGE. Gel Electrophoresis and Electroblotting of Proteins—Collagen fractions recovered by differential salt precipitation and column chromatography were routinely examined by SDS-PAGE (14Laemmli U.K. Nature. 1970; 227: 680-685Google Scholar). For amino-terminal sequence analysis and Western blotting, resolved protein bands were transblotted to PVDF membrane (Bio-Rad) using a MilliBlot-SDE electroblotting apparatus (Millipore). The membrane was washed thoroughly in ultrapure water (Millipore Milli-Q) to remove salts, then stained with Coomassie Brilliant Blue to detect protein bands. Bands were excised for automated sequence analysis (5Wu J.-J. Woods P.E. Eyre D.R. J. Biol. Chem. 1992; 267: 23007-23014Google Scholar). For Western blotting, a monoclonal antibody, 10F2, one of several raised against protease-generated neoepitopes in the collagen α1(II)C-telopeptide (15Atley L. Shao P. Ochs V. Shaffer K. Eyre D.R. Trans. Orthop. Res. Soc. 1998; 23: 850Google Scholar), was used as the primary, and biotin-conjugated goat anti-mouse IgG as the secondary, antibody then followed by ExtrAvidin-alkaline phosphatase (Sigma). The blots were developed using 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium. Protein Sequencing—Amino-terminal sequence analysis was performed on a Porton 2090E gas-phase microsequencer equipped with on-line high performance liquid chromatography analysis of phenylthiohydantoin derivatives. To remove pyroglutamate from the NH2-terminal end of blocked collagen chains, protein was solubilized in 0.1 m sodium phosphate buffer, 0.01 m EDTA, 5 mm dithiothreitol, 5% (v/v) glycerol, pH 8.0, and treated with the enzyme pyroglutamate aminopeptidase (Roche Applied Science) at 37 °C for 5 h before SDS-PAGE and transblotting (16Wu J.-J. Eyre D.R. J. Biol. Chem. 1995; 270: 18865-18870Google Scholar). The 2.0 m NaCl-precipitated fraction of the pepsin digest of bovine nucleus pulposus was enriched in the triple-helical COL1 and COL2 domains of type IX collagen. The 4.0 m NaCl-precipitated fraction contained the COL3 domain (Fig. 1a), which was further purified by molecular sieve chromatography on a Bio-Gel A5m column (Fig. 1b). After identification by SDS-PAGE, fractions combining the individual COL3 domains (bar in Fig. 1b) were pooled and the chains resolved by reverse-phase HPLC on a C8 column. Fig. 2 shows the HPLC profile and SDS-PAGE resolution of the individual COL3(IX) chains. The protein bands were transblotted to PVDF membrane, and their identities established by amino-terminal sequence analysis. Fig. 2 shows the determined NH2-terminal sequences of the three gene products after treatment with pyroglutamine aminopeptidase to remove blocking pyroglutamate residues. Without pyroglutamine aminopeptidase treatment, all three component chains were NH2-terminally blocked.Fig. 2Reverse-phase HPLC of type IX collagen COL3 domain from nucleus pulposus. Fractions marked by the bar in Fig. 1b were concentrated and eluted from a C8 reverse-phase column (Brownlee Aquapore RP-300; 4.6 mm × 25 cm) with a linear gradient (15–40%) of solvent B in A over 50 min at a flow rate of 1 ml/min collecting 1-ml fractions. Solvent A was 0.1% trifluoroacetic acid (v/v) in water, and solvent B was 0.085% trifluoroacetic acid (v/v) in acetonitrile: n-propyl alcohol (3:1, v/v). Aliquots of collected fractions (100 μl each) were analyzed by SDS-12.5% PAGE (inset). Also shown are amino-terminal sequences from the transblotted bands. Sequences were determined after removing the blocking pyroglutamate residue (Gln (Q)) with pyroglutamate aminopeptidase. P*, 4-hydroxyproline.View Large Image Figure ViewerDownload (PPT) When the α1(IX) NH2-terminal sequence derived from bovine nucleus pulposus was compared with the cDNA-predicted human sequence, nucleus pulposus α1(IX) was identical to the predicted short-form of human α1(IX) beginning with an NH2-terminal glutamate encoded by alternative exon 1 and continuing with the sequence encoded by exon 8 (Fig. 3a). This result establishes that bovine nucleus pulposus contains the short form of α1(IX) that lacks the NC4 protein domain. In contrast, NH2-terminal sequence analysis of pepsin-derived α1(IX)COL3 from calf articular cartilage showed that the amino terminus was not blocked and the sequence was identical to that of the long form of α1(IX) predicted from the human cDNA corresponding to the COOH-terminal portion of the NC4 domain (Fig. 3b). The results, however, show that the 7-amino acid sequence encoded by a predicted exon 7 is missing. The present findings, therefore, suggest a bovine splicing variant (or genomic difference) that results in the long α1(IX) form of bovine hyaline cartilage lacking the exon 7 sequence of the NC4 domain. Previous studies have shown that all four triple-helical cross-linking sites of type IX collagen are located in the COL2 domain (5Wu J.-J. Woods P.E. Eyre D.R. J. Biol. Chem. 1992; 267: 23007-23014Google Scholar, 17Diab M. Wu J.-J. Eyre D.R. Biochem. J. 1996; 314: 327-332Google Scholar). To determine whether the apparent exclusive usage of the short form of α1(IX) in the nucleus pulposus affects the cross-linking properties of type IX collagen, the 2 m NaCl-precipitated fraction of a pepsin digest of bovine nucleus pulposus was run on a C8 reverse-phase column to separate the COL2(IX) trimer from the COL1(IX) trimer (Fig. 4). Individual COL2(IX) chains were resolved from each other by reverse-phase HPLC (Fig. 5a) and SDS-PAGE (Fig. 5b) after reduction and alkylation of cysteine residues. Direct NH2-terminal sequence analysis of each protein band was performed after electroblotting to a PVDF membrane. Fig. 5d shows the sequences identified. Each type IX band gave multiple running sequences, one of which was the NH2 terminus of the NC3/COL2 domain. The others were the N- and C-telopeptides of type II collagen and α3(IX) NC1. All the results were consistent with previous findings on cartilage type IX collagen (5Wu J.-J. Woods P.E. Eyre D.R. J. Biol. Chem. 1992; 267: 23007-23014Google Scholar, 17Diab M. Wu J.-J. Eyre D.R. Biochem. J. 1996; 314: 327-332Google Scholar), with an N-telopeptide cross-linked to all three type IX chains at a COL2 site, α3(IX) NC1 linked to α1(IX) and α3(IX) at this same COL2 site and a C-telopeptide of type II collagen linked to a second α3(IX)COL2 site.Fig. 5Reverse-phase HPLC fractionation of individual collagen IX COL2 chains. Cysteine residues were reduced and carboxymethylated prior to chromatography on C8 reverse-phase HPLC with a linear gradient (15–40%) of solvent B in A over 50 min. a, reverse-phase HPLC profile. b, SDS-7.5% PAGE of HPLC-resolved type IX collagen COL2 domains stained with Coomassie Brilliant Blue. c, Western blot analysis of a duplicate gel using a monoclonal antibody that recognized the α1(II) C-telopeptide. Only the α3(IX)-COL2 domain reacted with the antibody. d, results of direct NH2-terminal sequencing of the electroblotted bands showing the individual α1(IX)-, α2(IX)-, and α3(IX)COL2 sequences and the peptides cross-linked to these COL2 helical domains. P*, 4-hydroxyproline; X, cross-linking hydroxylysine residue.View Large Image Figure ViewerDownload (PPT) Cross-linking between the type II collagen C-telopeptide and α3(IX)COL2 was further confirmed using mAb 10F2 (Fig. 5c), which recognizes a proteolytic neoepitope in the type II collagen C-telopeptide sequence, KGPDPLQ, in which the COOH-terminal glutamine is essential for immunoreactivity. The results of analysis of the COL3 domain of type IX collagen from nucleus pulposus establish that only the short form of α1(IX) (expressed by use of alternative exon 1) is used in the matrix. Several other collagen genes have been reported to use alternative promoters and produce alternative transcripts in a tissue-specific manner. For example, with human α2(VI) and mouse α1(XVIII) genes, the translated products of two different promoters vary in the length of their NH2-terminal non-collagenous domain (18Saitta B. Chu M.L. Eur. J. Biochem. 1994; 223: 675-682Google Scholar, 19Rehn M. Hintikka E. Pihlajaniemi T. Genomics. 1996; 32: 436-446Google Scholar). From chick α2(I) and α1(III) genes, the alternative transcripts create different reading frames that do not encode collagenous proteins (20Pallante K.M. Niu Z. Zhao Y. Cohen A.J. Nah H.D. Adams S.L. J. Biol. Chem. 1996; 271: 25233-25239Google Scholar, 21Zhang Y. Niu Z. Cohen A.J. Nah H.D. Adams S.L. Nucleic Acids Res. 1997; 25: 2470-2477Google Scholar, 22Cohen A.J. Lakshmi T.R. Niu Z. Trindade J. Billings P.C. Adams S.L. Mech. Dev. 2002; 114: 177-180Google Scholar). The short transcriptional form of α1(IX) has been identified in the developing chick cornea, vitreous humor, and notochord and in mouse eye and heart during embryonic development (12Nishimura I. Muragaki Y. Olsen B.R. J. Biol. Chem. 1989; 264: 20033-20041Google Scholar, 23Yada T. Suzuki S. Kobayashi K. Kobayashi M. Hoshino T. Horie K. Kimata K. J. Biol. Chem. 1990; 265: 6992-6999Google Scholar, 24Brewton R.G. Wright D.W. Mayne R. J. Biol. Chem. 1991; 266: 4752-4757Google Scholar, 25Hayashi M. Hayashi K. Iyama K. Trelstad R.L. Linsenmayer T.F. Mayne R. Dev. Dyn. 1992; 194: 169-176Google Scholar, 26Swiderski R.E. Solursh M. Dev. Dyn. 1992; 194: 118-127Google Scholar, 27Liu C.-Y. Olsen B.R. Kao W.W.-Y. Dev. Dyn. 1993; 198: 150-157Google Scholar, 28Fitch J.M. Gordon M.K. Gibney E.P. Linsenmayer T.F. Dev. Dyn. 1995; 202: 42-53Google Scholar). In the young disc, nucleus pulposus is known to contain both chondrocytic and notochordal cells (29Oegema Jr., T.R. Biochem. Soc. Trans. 2002; 30: 839-844Google Scholar). During growth, the notochordal cells die while chondrocytic cells persist. Since only one transcription product, the short form of α1(IX), was detected in the young bovine nucleus pulposus, this suggests that either the chondrocytic cells of the nucleus differ from those of hyaline cartilage in their promoter usage of Col9a1 or, perhaps, that the notochordal cells produce the collagen IX. It is notable in this regard that in animals in which the nucleus pulposus retains a gelatinous, youthful texture throughout life (e.g. non-chondrodystrophoid breeds of dog), notochordal cells also persist (29Oegema Jr., T.R. Biochem. Soc. Trans. 2002; 30: 839-844Google Scholar). The short form of the type IX collagen molecule in chick vitreous also bears a long glycosaminoglycan chain on the NC3 domain of the α2(IX) chain (23Yada T. Suzuki S. Kobayashi K. Kobayashi M. Hoshino T. Horie K. Kimata K. J. Biol. Chem. 1990; 265: 6992-6999Google Scholar). The functional significance of the long glycosaminoglycan chain is still uncertain, although it has been proposed that it might prevent lateral fibril aggregation in the gel-like vitreous. Collagen fibrils in the vitreous are particularly thin and widely dispersed from each other compared with cartilage (30Bos K.J. Holmes D.F. Kadler K.E. McLeod D. Morris N.P. Bishop P.N. J. Mol. Biol. 2001; 306: 1011-1022Google Scholar), in which in the mammal, the α2(IX) chain lacks the glycosaminoglycan chain (17Diab M. Wu J.-J. Eyre D.R. Biochem. J. 1996; 314: 327-332Google Scholar). Thin fibrils, and a gelatinous texture, are also features of the young nucleus pulposus. It is not known whether the α2(IX) chain of nucleus pulposus has a glycosaminoglycan chain attached to its NC3 domain. The present findings do show that despite lacking an NC4 domain on the α1(IX) chain, the cross-linking properties of nucleus pulposus type IX collagen are essentially the same as in cartilage. As in cartilage, the disc-type IX molecules are cross-linked to type II collagen and to other type IX collagen molecules, so that a recently proposed model of the molecular interactions (31Eyre D.R. Wu J.-J. Fernandes R.J. Pietka T.A. Weis M.A. Biochem. Soc. Trans. 2002; 30: 894-899Google Scholar), which accommodates all known cross-linking sites, is applicable to disc and cartilage. The results also show that the 7-amino acid sequence encoded by human exon 7 is absent from bovine α1(IX). It has been established for chick cartilage that the equivalent exon 7 sequence is present in the cDNA from the long mRNA transcript of Col9a1 and by sequence analysis of tryptic peptides that the protein sequence is expressed in tissue α1(IX) (7Vasios G. Nishimura I. Konomi H. van der Rest M. Ninomiya Y. Olsen B.R. J. Biol. Chem. 1988; 263: 2324-2329Google Scholar). The present findings, therefore, indicate either a gene sequence difference in bovine Col9a1 or a spliced-out version of α1(IX) lacking the exon 7 sequence in our bovine cartilage samples. No evidence emerged for an α1(IX) chain that included the exon 7 domain, so it is not known whether the alternative version can be expressed in any tissue in this species. Bovine genomic sequence is not available to help resolve this issue. The α1(IX) chain NC4 domain is encoded by exons 2–7. It is not known whether any exons other than 7 can be variably spliced. Transcripts of the other cartilage-specific collagens, types II and XI, are known to be variably spliced (32Ryan M.C. Sandell L.J. J. Biol. Chem. 1990; 265: 10334-10339Google Scholar, 33Oxford J.T. Doege K.J. Morris N.P. J. Biol. Chem. 1995; 270: 9478-9485Google Scholar, 34Zhidkova N.I. Justice S.K. Mayne R. J. Biol. Chem. 1995; 270: 9486-9493Google Scholar, 35Zhu Y. McAlinden A. Sandell L.J. Dev. Dyn. 2001; 220: 350-362Google Scholar). Tryptophan substitutions at position 326 of the α2(IX) chain, and position 103 of the α3(IX) chain, have been linked to increased risk of human lumbar disc degeneration and chronic sciatica (8Annunen S. Paassilta P. Lohiniva J. Perala M. Pihlajamaa T. Karppinen J. Tervonen O. Kroger H. Lahde S. Vanharanta H. Ryhanen L. Goring H.H.H. Ott J. Prockop D.J. Ala-Kokko L. Science. 1999; 285: 409-412Google Scholar, 9Paassilta P. Lohiniva J. Goring H.H.H. Perala M. Raina S.S. Karppinen J. Hakala M. Palm T. Kroger H. Kaitila I. Vanharanta H. Ott J. Ala-Kokko L. J. Am. Med. Assoc. 2001; 285: 1843-1849Google Scholar). It has been proposed that the tryptophan residue in the α2(IX)COL2 domain near the known cross-linking site at residue 330 in α3(IX)COL2 might interfere with the interaction between collagens II and IX (8Annunen S. Paassilta P. Lohiniva J. Perala M. Pihlajamaa T. Karppinen J. Tervonen O. Kroger H. Lahde S. Vanharanta H. Ryhanen L. Goring H.H.H. Ott J. Prockop D.J. Ala-Kokko L. Science. 1999; 285: 409-412Google Scholar). This site cross-links the C-telopeptide of type II collagen to α3(IX)COL2. Western blot analysis using monoclonal antibody 10F2, which is specific to the C-telopeptide of type II collagen, confirms that the type II collagen C-telopeptide is attached selectively to α3(IX)COL2 (Fig. 5c). Antibody 10F2 recognizes a cleavage site in the type II C-telopeptide generated by pepsin. 2J.-J. Wu and D. R. Eyre, unpublished observations. The yields on NH2-terminal sequence analysis also indicate that essentially every molecule has an attached C-telopeptide. Future work with this antibody might, therefore, provide a sensitive method for detecting any changes in cross-linking between α3(IX)COL2 and type II collagen C-telopeptide caused by the tryptophan-containing polymorphic variants (38Matsui Y. Wu J.-J. Weis M.A. Pietka T. Eyre D.R. Matrix Biol. 2003; 22: 123-129Google Scholar). Peptide mapping using a multi-epitope recognizing antiserum to type IX collagen has proved to be useful for displaying the cross-linking properties in general of type IX collagen in fetal human cartilage (36Ichimura S. Wu J.-J. Eyre D.R. Arch. Biochem. Biophys. 2000; 378: 33-39Google Scholar, 37Bönnemann C.G. Cox G.F. Shapiro F. Wu J.-J. Feener C.A. Thompson T.G. Anthony D.C. Eyre D.R. Darras B.T. Kunkel L.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1212-1217Google Scholar, 38Matsui Y. Wu J.-J. Weis M.A. Pietka T. Eyre D.R. Matrix Biol. 2003; 22: 123-129Google Scholar). In conclusion, we have found that nucleus pulposus of the intervertebral disc contains the short form of the α1(IX) chain, a product of an alternative promoter, which lacks the NC4 domain. The cross-linking properties of nucleus pulposus type IX collagen are, however, similar to those of hyaline cartilage. We also have identified in bovine cartilage a previously undescribed splicing variant of the long form of α1(IX) that lacks the sequence encoded by exon 7. Its pattern of expression (in development, tissue type, species, etc.), and any functional significance remains unknown." @default.
• W2000783985 created "2016-06-24" @default.
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• W2000783985 date "2003-07-01" @default.
• W2000783985 modified "2023-09-30" @default.
• W2000783985 title "Intervertebral Disc Collagen" @default.
• W2000783985 cites W1486071978 @default.
• W2000783985 cites W1505294312 @default.
• W2000783985 cites W1515000724 @default.
• W2000783985 cites W1525176931 @default.
• W2000783985 cites W1525977186 @default.
• W2000783985 cites W1526955320 @default.
• W2000783985 cites W1528604920 @default.
• W2000783985 cites W1542740939 @default.
• W2000783985 cites W1555090655 @default.
• W2000783985 cites W1557792852 @default.
• W2000783985 cites W1565114828 @default.
• W2000783985 cites W166713157 @default.
• W2000783985 cites W1964082546 @default.
• W2000783985 cites W1968817960 @default.
• W2000783985 cites W1973547300 @default.
• W2000783985 cites W1976429655 @default.
• W2000783985 cites W1976763134 @default.
• W2000783985 cites W1999865365 @default.
• W2000783985 cites W2015712262 @default.
• W2000783985 cites W2016283013 @default.
• W2000783985 cites W2019613296 @default.
• W2000783985 cites W2046898502 @default.
• W2000783985 cites W2067283439 @default.
• W2000783985 cites W2075488484 @default.
• W2000783985 cites W2078744277 @default.
• W2000783985 cites W2079047427 @default.
• W2000783985 cites W2080865520 @default.
• W2000783985 cites W2088434470 @default.
• W2000783985 cites W2089315153 @default.
• W2000783985 cites W2089394770 @default.
• W2000783985 cites W2092708316 @default.
• W2000783985 cites W2098102950 @default.
• W2000783985 cites W2100837269 @default.
• W2000783985 cites W2129465868 @default.
• W2000783985 cites W2142182803 @default.
• W2000783985 cites W2172259165 @default.
• W2000783985 cites W4246933005 @default.
• W2000783985 doi "https://doi.org/10.1074/jbc.m302431200" @default.
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