Identification of a stromelysin cleavage site within the interglobular domain of human aggrecan. Evidence for proteolysis at this site in vivo in human articular cartilage.
Products generated by the digestion of human aggrecan with recombinant human stromelysin have been purified and analyzed by N-terminal sequencing and C-terminal peptide isolation. N-terminal analysis of chondroitin sulfate-bearing fragments revealed a clearly identifiable sequence initiating at residue Phe342 of human aggrecan, providing evidence for a cleavage site at the Asn341-Phe342 bond located within the interglobular domain. This cleavage site, which separates the G1 domain from the remainder of the molecule, was confirmed by isolation from the liberated G1 domain of a C-terminal tryptic peptide with the sequence YDAICYTGEDFVDIPEN (in which the C-terminal residue is Asn341). This peptide was also isolated from tryptic digests of hyaluronan-binding proteins (A1D4 samples) prepared by CsCl gradient centrifugation of extracts of mature human articular cartilages. Since these A1D4 samples contain G1 domain which accumulates as a result of aggrecan catabolism in vivo, these results clearly indicate that stromelysin cleaves the Asn341-Phe342 bond of human aggrecan in situ.
Flannery
C R
CR
Shriners Hospitals for Crippled Children, Tampa, Florida 33612.
Lark
M W
MW
Sandy
J D
JD
D000368:Aged; D053674:Aggrecans; D000375:Aging / Q000378:metabolism; D000595:Amino Acid Sequence; D002356:Cartilage / Q000378:metabolism*; D002809:Chondroitin Sulfates / Q000378:metabolism; D016326:Extracellular Matrix Proteins; D005260:Female; D006801:Humans; D006868:Hydrolysis; D037181:Lectins, C-Type; D008297:Male; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D008875:Middle Aged; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism*; D014357:Trypsin / Q000737:chemistry
D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D002809:Chondroitin Sulfates; D014357:Trypsin; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3