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PubMed · 7806222

Complete coding sequence, deduced primary structure, chromosomal localization, and structural analysis of murine aggrecan.

E Walcz·F Deák·P Erhardt·S N Coulter·C Fülöp·P Horvath·K J Doege·T T Glant
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Abstract

We have isolated and sequenced overlapping cDNA clones encoding the entire core protein of aggrecan (the large aggregating chondroitin sulfate/keratan sulfate proteoglycan of cartilage) from three chondrocyte cDNA libraries of BALB/c mice and localized the aggrecan gene in mouse chromosome 7. We determined 7386 bp of the cDNA sequence, including 132 and 854 nucleotides of 5' and 3' untranslated regions, respectively. The core protein precursor is 2132 amino acids long (M(r) 222,008), including a 19-residue secretory signal peptide. The overall amino acid sequence of the mouse aggrecan shows 91.6% identity to rat and 72.5% to human aggrecan. Comparison of the amino acid sequences of various domains and subdomain structures of mouse aggrecan to known sequences of other species and related proteins (versican, neurocan, link protein, and lymphocyte homing receptor CD44) revealed high levels of identity of the G1, G2, and G3 globular domains and relatively less conserved structures in the interglobular and glycosaminoglycan-attachment regions. Epidermal growth factor (EGF)-like module was detected in only a minor fraction of aggrecan clones, while the complement regulatory protein (CRP)-like domain was regularly expressed in all samples.

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AggrecansAmino Acid SequenceAnimalsBase SequenceChromosome MappingDNA, ComplementaryExtracellular Matrix ProteinsGene ExpressionGene LibraryHumansLectins, C-TypeMiceMice, Inbred StrainsMolecular Sequence DataProtein PrecursorsProteoglycansRatsSequence Homology, Amino AcidSpecies SpecificityComparative StudyJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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E Walcz, F Deák, P Erhardt, S N Coulter, C Fülöp, P Horvath, K J Doege, T T Glant. 1994-07-15. Complete coding sequence, deduced primary structure, chromosomal localization, and structural analysis of murine aggrecan.. https://doi.org/10.1006/geno.1994.1396

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PubMed1995

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Brevican is a member of the aggrecan family of chondroitin sulfate proteoglycans that is predominantly expressed in the nervous system. In the adult brain, the brevican core protein undergoes proteolytic cleavage and exists as a 145-kDa full-length form and an 80-kDa C-terminal fragment. We have determined the complete primary structure of rat brevican and the N-terminal amino acid sequence of the 80-kDa fragment. The results demonstrate that the cleavage of the brevican core protein occurs in a highly conserved region within a generally poorly conserved central domain, and that the sequence at the cleavage site shows intriguing similarities to the "aggrecanase" cleavage site in aggrecan. cDNA cloning of rat brevican has also revealed that the putative hyaluronic acid binding protein, BEHAB, is identical to the N-terminal half of brevican.

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PubMed1995

Neuroglycan C, a novel membrane-spanning chondroitin sulfate proteoglycan that is restricted to the brain.

Monoclonal antibodies were raised to membrane-bound proteoglycans derived from rat brain, and four monoclonal antibodies that recognized a 150-kDa chondroitin sulfate proteoglycan with a core glycoprotein of 120 kDa were obtained. Immunohistological study revealed that the proteoglycan was associated with developing neurons. We screened rat brain cDNA libraries using the four monoclonal antibodies and isolated overlapping cDNA clones that encoded the entire core protein of 514 amino acids plus a 30-residue signal peptide. The deduced amino acid sequence suggested an integral membrane protein divided into five structurally different domains: an N-terminal domain to which chondroitin sulfate chains might be attached, a basic amino acid cluster consisting of seven arginine and two lysine residues, a cysteine-containing domain, a membrane-spanning segment, and a C-terminal cytoplasmic domain of 95 amino acids. On Northern blots, the cDNA hybridized with a single mRNA of 3.1 kilobases that was detectable in brains of neonatal and adult rats but not in kidney, liver, lung, and muscle of either. The sequence of the proteoglycan did not exhibit significant homology to any other known protein, indicating that the proteoglycan, designated neuroglycan C, is a novel integral membrane proteoglycan.

Aggrecans↗
PubMed1995

Catabolism of aggrecan by explant cultures of human articular cartilage in the presence of retinoic acid.

The N-terminal amino acid sequence of human aggrecan was determined and it was shown that two sequences were present. The major sequence, AVTVE-, accounted for 60% of the aggrecan and started at alanine residue 17 of the human aggrecan core protein cDNA sequence (K. Doege et al. (1991) J. Biol. Chem. 266, 894-920). The other N-terminal sequence, VETX-, started at valine residue 20. Characterization of aggrecan core protein peptides present in the matrix of adult human articular cartilage showed that at least 11 aggrecan core proteins were present with approximate M(r) between 300,000 and 43,000. All these core proteins were found to have the same N-terminal sequences as that observed in human aggrecan. When articular cartilage was placed in explant culture in medium containing 10(-6) M retinoic acid there was a 3.5-fold increase in the loss of aggrecan into the culture medium compared to tissue maintained in medium alone or medium containing 20% (v/v) newborn calf serum. Analysis of the aggrecan core protein fragments that were released to the culture medium containing 10(-6) M retinoic acid showed the presence of 13 core protein peptides of M(r) between 300,000 and 43,000. The 11 smaller peptides of M(r) 230,000 to 43,000 were shown to have the N-terminal sequence ARGS-. This sequence which starts at residue 393 of the human aggrecan core protein is located within the interglobular region between the G1 and G2 domains and is the site of aggrecan catabolism by the putative protease aggrecanase. The presence of core proteins of varying sizes but with the same N-terminal sequence reflects proteolytic processing from the C-terminal end of the core protein that was also observed in the aggrecan macromolecules extracted from the matrix of human articular cartilage. This proteolytic processing was also evident but to a lesser extent in newly synthesized 35S-labeled aggrecan macromolecules.

Aggrecans↗