Complete Coding Sequence, Deduced Primary Structure, Chromosomal Localization, and Structural Analysis of Murine Aggrecan

Walcz, E; Deák, F.; Erhardt, Péter; Coulter, Silvija N.; Fülöp, Csaba; Horváth, Péter; Doege, Kurt; Glant, Tibor T.

doi:10.1006/geno.1994.1396

Cited by 56 publications

(44 citation statements)

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“…3A). These aggrecanase-specific cleavage sites have been confirmed in mouse aggrecan with antibodies to neoepitopes that result from the cleavage of the core protein at Glu 373 -Ala, Glu 1279 -Gly, Glu 1467 -Gly, and Glu 1572 -Ala bonds (5,9,22,34,35,42).…”

Section: Rate Of Loss Ofmentioning

confidence: 79%

“…This is not surprising given the similarity of susceptible sites along the core protein and the carboxyl terminally driven process of aggrecan degradation in cartilage (43). The homologous cleavage sites in mouse aggrecan (42) are located at Glu 373 -Ala within the interglobular domain, at Glu 1279 -Gly between the CS-1 and CS-2 domains, and at Glu 1467 -Gly, Glu 1572 -Ala, and Glu 1672 -Leu bonds within the CS-2 domain (Fig. 3A).…”

Section: Rate Of Loss Ofmentioning

confidence: 97%

“…In IL-1␣-treated tissue, significant levels of peptide 2b and peptides 3, 4, and 6 appeared in the matrix and medium, respectively, whereas there was only a marginal effect on the levels of peptides 5 and 7 (Fig. 2D) Western Blot Analysis of Aggrecan Fragments-To monitor the degree of proteolytic processing of total aggrecan and to correlate this with the radiolabeled fragments, aliquots of media and extracts were fractionated on SDS gels and analyzed by Western blotting with neoepitope antibodies (9,22,35,42). The immunopositive bands were matched to bands detected on the same membrane with antibody 2B6 (41) to show the overall pattern of degradation products.…”

Section: Rate Of Loss Ofmentioning

confidence: 99%

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Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

Ilic

East

Rogerson

et al. 2007

Journal of Biological Chemistry

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Aggrecan loss from mouse cartilage is predominantly because of ADAMTS-5 activity; however, the relative contribution of other proteolytic and nonproteolytic processes to this loss is not clear. This is the first study to compare aggrecan loss with aggrecan processing in mice with single and double deletions of ADAMTS-4 and -5 activity (⌬cat). Cartilage explants harvested from single and double ADAMTS-4 and -5 ⌬cat mice were cultured with or without interleukin (IL)-1␣ or retinoic acid and analyzed for (i) the kinetics of 35 S-labeled aggrecan loss, (ii) the pattern of 35 S-labeled aggrecan fragments released into the media and retained in the matrix, (iii) the pattern of total aggrecan fragments released into the media and retained in the matrix, and (iv) specific cleavage sites within the interglobular and chondroitin sulfate-2 domains. The loss of radiolabeled aggrecan from ADAMTS-4/-5 ⌬cat cartilage was less than that from ADAMTS-4, ADAMTS-5, or wild-type cartilage under nonstimulated conditions. IL-1␣ and retinoic acid stimulated radiolabeled aggrecan loss from wild-type and ADAMTS-4 ⌬cat cartilage, but there was little effect on ADAMTS-5 cartilage. Proteolysis of aggrecan contributed most to its loss in wild-type, ADAMTS-4, and ADAMTS-5 ⌬cat cartilage explants. The pattern of proteolytic processing of aggrecan in these cultures was consistent with that occurring in cartilage pathologies. Retinoic acid, but not IL-1␣, stimulated radiolabeled aggrecan loss from ADAMTS-4/-5 ⌬cat cartilage explants. Even though there was a 300% increase in aggrecan loss from ADAMTS-4/-5 ⌬cat cartilage stimulated with retinoic acid, the loss was not associated with aggrecanase cleavage but with the release of predominantly intact aggrecan consistent with the phenotype of the ADAMTS-4/-5 ⌬cat mouse. Our results show that chondrocytes have additional mechanism for the turnover of aggrecan and that when proteolytic mechanisms are blocked by ablation of aggrecanase activity, nonproteolytic mechanisms compensate to maintain cartilage homeostasis.Aggrecan catabolism has been studied extensively in the past 30 -40 years as part of the metabolic processes important in normal functional cartilage and in cartilage pathology. The major enzymes involved in degradation of aggrecan in normal and pathological cartilage are aggrecanases members of the ADAMTS 2 subfamily of adamalysin (M12) metalloproteinases (1, 2). These enzymes cleave the aggrecan core protein at aggrecanase-specific cleavage sites between glutamate and a small hydrophobic amino acid residue (3, 4). Most studies suggest that in vitro and in vivo ADAMTS-4 and -5 are the main aggrecanases (5-11); however, ADAMTS-1, -8, -9, -15, -16, and -18 also exhibit aggrecanase activity (12-15). Recombinant ADAMTS-4 and -5 have been suggested to be the most potent aggrecanases (15-19), although the relative potency in aggrecan degrading activity has yet to be clarified (11,20,21). In addition, recombinant ADAMTS-1, -4, and -5 cleave aggrecan at 5 sites: one within the interglobular domain a...

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Section: Rate Of Loss Ofmentioning

confidence: 79%

Section: Rate Of Loss Ofmentioning

confidence: 97%

Section: Rate Of Loss Ofmentioning

confidence: 99%

See 1 more Smart Citation

Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

Ilic

East

Rogerson

et al. 2007

Journal of Biological Chemistry

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“…The rat (5), mouse (16), and chicken (17) aggrecan cDNA sequences have been reported, and although a recognizable variant of the human repeat region can be seen in all cases, the sequences are poorly conserved as a group for each species relative to the level of the human repeats. Partial bovine cDNA sequences have been published (13,14), but these non-overlapping clones do not include the CS domain sequence where the repeat region would be expected.…”

Section: Resultsmentioning

confidence: 99%

A Human-specific Polymorphism in the Coding Region of the Aggrecan Gene

Doege

Coulter

Meek

et al. 1997

Journal of Biological Chemistry

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111

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“…It is a functionally important component of both articular cartilage [1] and intervertebral disc [2]. Cloning and sequencing of the aggrecan cDNA and gene from different species have allowed an extensive characterization of the molecule at the structural level [3][4][5]. Aggrecan consists of a core protein with several distinct domains.…”

Section: Introductionmentioning

confidence: 99%

Aggrecan degradation in human intervertebral disc and articular cartilage

Sztrolovics

Alini

Roughley

et al. 1997

Biochemical Journal

242

172

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Aggrecan degradation in human intervertebral disc and articular cartilage has been studied by using anti-neoepitope antibodies specific for the N-terminal degradation products generated by cleavage within the interglobular domain at the metalloproteinase and aggrecanase sites. Immunoblot analysis of extracts of annulus fibrosus, nucleus pulposus and articular cartilage demonstrated age-related patterns in the abundance of both degradation products. In all three tissues the metalloproteinase-generated fragment was present at very low levels in young individuals but increased in abundance with age. In the disc tissues, the abundance of this degradation product levelled off in the juvenile; for cartilage this occurred in early adulthood. Despite these temporal differences, the levels attained in adults were comparable for the three tissues. In contrast, the aggrecanase-generated degradation product exhibited tissue-specific differences in the variation of its abundance with age. Whereas this degradation product increased with age in annulus fibrosus and articular cartilage and had levelled off by adulthood, in nucleus pulposus it was present in greatest abundance in young individuals and decreased to very low levels with age. Examination of discs exhibiting various degrees of degeneration did not reveal any differences in the levels of the metalloproteinase and aggrecanase-generated cleavage products that could not be accounted for by differences in age. In adults the product of aggrecanase action was much more abundant in articular cartilage than in either of the disc tissues, despite the age-related increase also observed for annulus fibrosus. Analysis of tissue extracts with an antibody recognizing the G1 domain of aggrecan identified two major degradation products whose abundance and size were correlated with the fragments detected by the anti-neoepitope antibodies. Taken together, these results indicate that cleavage at the metalloproteinase and aggrecanase sites are quantitatively important events in aggrecan catabolism in both articular cartilage and intervertebral disc in vivo. Moreover the two enzyme systems act independently and exhibit differences in the degree to which they contribute to aggrecan degradation in these tissues.

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Complete Coding Sequence, Deduced Primary Structure, Chromosomal Localization, and Structural Analysis of Murine Aggrecan

Cited by 56 publications

References 0 publications

Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

A Human-specific Polymorphism in the Coding Region of the Aggrecan Gene

Aggrecan degradation in human intervertebral disc and articular cartilage

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