The fibril-associated collagen IX provides a novel mechanism for cell adhesion to cartilaginous matrix.
Collagen IX is the prototype fibril-associated collagen with interruptions in triple helix. In human cartilage it covers collagen fibrils, but its putative cellular receptors have been unknown. The reverse transcription-PCR analysis of human fetal tissues suggested that based on their distribution all four collagen receptor integrins, namely alpha1beta1, alpha2beta1, alpha10beta1, and alpha11beta1, are possible receptors for collagen IX. Furthermore primary chondrocytes and chondrosarcoma cells express the four integrins simultaneously. Chondrosarcoma cells, as well as Chinese hamster ovary cells transfected to express alpha1beta1, alpha2beta1, or alpha10beta1 integrin as their only collagen receptor, showed fast attachment and spreading on human recombinant collagen IX indicating that it is an effective cell adhesion protein. To further study the recognition of collagen IX we produced recombinant alphaI domains in Escherichia coli. For each of the four alphaI domains, collagen IX was among the best collagenous ligands, making collagen IX exceptional compared with all other collagen subtypes tested so far. Rotary shadowing electron microscopy images of both alpha1I- and alpha2I-collagen IX complexes unveiled only one binding site located in the COL3 domain close to the kink between it and the COL2 domain. The recognition of collagen IX by alpha2I was considered to represent a novel mechanism for two reasons. First, collagen IX has no GFOGER motif, and the identified binding region lacks any similar sequences. Second, the alpha2I domain mutations D219R and H258V, which both decreased binding to collagen I and GFOGER, had very different effects on its binding to collagen IX. D219R had no effect, and H258V prevented type IX binding. Thus, our results indicate that collagen IX has unique cell adhesion properties when compared with other collagens, and it provides a novel mechanism for cell adhesion to cartilaginous matrix.
Käpylä
Jarmo
J
Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylä FI-40014, Finland.
Jäälinoja
Juha
J
Tulla
Mira
M
Ylöstalo
Joni
J
Nissinen
Liisa
L
Viitasalo
Tiina
T
Vehviläinen
Piia
P
Marjomäki
Varpu
V
Nykvist
Petri
P
Säämänen
Anna-Marja
AM
Farndale
Richard W
RW
Birk
David E
DE
Ala-Kokko
Leena
L
Heino
Jyrki
J
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002356:Cartilage / Q000737:chemistry / Q000378:metabolism*; D002448:Cell Adhesion; D002460:Cell Line; D045744:Cell Line, Tumor; D019902:Chondrocytes / Q000378:metabolism; D002813:Chondrosarcoma / Q000378:metabolism; D003094:Collagen / Q000737:chemistry / Q000378:metabolism; D024081:Collagen Type IX / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006224:Cricetinae; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D039001:Integrin alpha Chains / Q000096:biosynthesis; D039423:Integrin alpha1 / Q000096:biosynthesis; D039421:Integrin alpha2 / Q000096:biosynthesis; D008024:Ligands; D051379:Mice; D008854:Microscopy, Electron; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D024081:Collagen Type IX; C121535:ITGA11 protein, human; D039001:Integrin alpha Chains; D039423:Integrin alpha1; D039421:Integrin alpha2; D008024:Ligands; D010455:Peptides; D012333:RNA, Messenger; D011994:Recombinant Proteins; C406210:integrin alpha 10; D012313:RNA; D003094:Collagen