Molecular properties of matrilin-3 isolated from human growth cartilage.
Matrilin-3 is a recently identified matrix protein of cartilage that shows sequence homology to matrilin-1 (cartilage matrix protein or CMP). Here we identify and characterize the molecular properties of matrilin-3 from human growth cartilage by immunochemical and mass spectrometry methods. Extracts of fetal skeletal cartilage were resolved by SDS-PAGE and candidate matrilin subunits were identified by electrospray mass spectrometry of tryptic peptides. Matrilin-3 and matrilin-1 were both present in disulfide-bonded tetrameric components. Polyclonal antisera to synthetic peptides specific to each subunit confirmed the identities by Western blotting and further demonstrated the existence of several forms of tetramer. A homotetramer (matrilin-3)4 and more than one species of heterotetramer containing matrilin-3 and matrilin-1 chains were resolved. Immunohistochemistry of tissue sections confirmed that both matrilin-1 and matrilin-3 are widely codistributed throughout human skeletal growth cartilage.
Kleemann-Fischer
D
D
Department of Orthopaedics, University of Washington, Seattle 98195, USA.
Kleemann
G R
GR
Engel
D
D
Yates
J R
JR
Wu
J J
JJ
Eyre
D R
DR
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D064236:Cartilage Oligomeric Matrix Protein; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism; D006132:Growth Plate / Q000737:chemistry* / Q000196:embryology / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D013058:Mass Spectrometry; D064235:Matrilin Proteins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D020539:Sequence Analysis, Protein
D064236:Cartilage Oligomeric Matrix Protein; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; C577539:MATN1 protein, human; D064235:Matrilin Proteins; D010446:Peptide Fragments; C577553:TSP5 protein, human