The 67-kD elastin/laminin-binding protein is related to an enzymatically inactive, alternatively spliced form of beta-galactosidase.
We and others have previously shown that a 67-kD cell surface elastin/laminin-binding protein (EBP) is responsible for cell adhesion to elastin and laminin and for mediating the process of elastin fiber assembly, but the nature of this protein was unknown. In this report we provide evidence that a 67-kD catalytically inactive form of beta-galactosidase produced by alternative splicing demonstrates immunological and functional similarity and sequence homology to the 67-kD EBP, suggesting that the two might be the same. Antibody prepared to a synthetic peptide, N-Ac-GSPSAQDEASPL, corresponding to a frame-shift-generated sequence unique to the alternatively spliced form of human beta-galactosidase, also recognized sheep EBP both on Western blotting and in aortic tissue. Furthermore, this synthetic peptide (S-GAL) binds to elastin and laminin, but not to fibronectin, collagen I, or collagen III. Moreover, both tropoelastin and laminin which bind to S-GAL peptide affinity columns can be specifically eluted from them with an excess of free S-GAL peptides. In addition, sequence homology among this splice variant of human beta-galactosidase, sheep EBP, and NH2-terminal sequences of some elastases suggests that these proteins share a common ligand-binding motif that has not been previously recognized.
Division of Cardiovascular Research, Hospital for Sick Children, Toronto, Ontario, Canada.
A
A
Hinek
M
M
Rabinovitch
F
F
Keeley
Y
Y
Okamura-Oho
J
J
Callahan
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta / Q000378:metabolism; D015153:Blotting, Western; D004549:Elastin / Q000378:metabolism*; D005333:Fetus; D014644:Genetic Variation; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D007797:Laminin; D010455:Peptides; D011956:Receptors, Cell Surface; C054901:elastin-binding proteins; D004549:Elastin; D001616:beta-Galactosidase