Polydom/SVEP1 is a ligand for integrin α9β1.
A variety of proteins, including tenascin-C and osteopontin, have been identified as ligands for integrin α9β1. However, their affinities for integrin α9β1 are apparently much lower than those of other integrins (e.g. α3β1, α5β1, and α8β1) for their specific ligands, leaving the possibility that physiological ligands for integrin α9β1 still remain unidentified. In this study, we found that polydom (also named SVEP1) mediates cell adhesion in an integrin α9β1-dependent manner and binds directly to recombinant integrin α9β1 with an affinity that far exceeds those of the known ligands. Using a series of recombinant polydom proteins with N-terminal deletions, we mapped the integrin-binding site to the 21st complement control protein domain. Alanine-scanning mutagenesis revealed that the EDDMMEVPY sequence (amino acids 2636-2644) in the 21st complement control protein domain was involved in the binding to integrin α9β1 and that Glu(2641) was the critical acidic residue for the integrin binding. The importance of this sequence was further confirmed by integrin binding inhibition assays using synthetic peptides. Immunohistochemical analyses of mouse embryonic tissues showed that polydom colocalized with integrin α9 in the stomach, intestine, and other organs. Furthermore, in situ integrin α9β1 binding assays using frozen mouse tissues showed that polydom accounts for most, but not all, of the integrin α9β1 ligands in tissues. Taken together, the present findings indicate that polydom is a hitherto unknown ligand for integrin α9β1 that functions as a physiological ligand in vivo.
Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
Ryoko
R
Sato-Nishiuchi
Itsuko
I
Nakano
Akio
A
Ozawa
Yuya
Y
Sato
Makiko
M
Takeichi
Daiji
D
Kiyozumi
Kiyoshi
K
Yamazaki
Teruo
T
Yasunaga
Sugiko
S
Futaki
Kiyotoshi
K
Sekiguchi
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D006801:Humans; D016023:Integrins / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D051379:Mice; D016296:Mutagenesis; D009928:Organ Specificity / Q000502:physiology; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion
D002135:Calcium-Binding Proteins; D015815:Cell Adhesion Molecules; D016023:Integrins; D008024:Ligands; C418635:Polydom protein, mouse; D011506:Proteins; C508516:SVEP1 protein, human; C113256:integrin alpha 9 beta 1