Tropoelastin interacts with cell-surface glycosaminoglycans via its COOH-terminal domain.
Using a biochemical and cell biological approach, we have identified a cell interaction site at the carboxyl terminus of tropoelastin. Cell interactions with the COOH-terminal sequence are not through the elastin-binding protein (EBP67) because neither VGVAPG-like peptides nor galactoside sugars altered adhesion. Our results also show that cell adhesion to tropoelastin is not promoted by integrins. Through the use of mutant Chinese hamster ovary cell lines defective in glycosaminoglycan biosynthesis, as well as competition studies and enzymatic removal of specific cell-surface glycosaminoglycans, the tropoelastin-binding moieties on the cell surface were identified as heparan and chondroitin sulfate-containing glycosaminoglycans, with heparan sulfate being greatly preferred. Heparin affinity chromatography combined with cell adhesion assays identified the last 17 amino acids as the sequence element at the carboxyl terminus of tropoelastin responsible for the adhesive activity.
Broekelmann
Thomas J
TJ
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Kozel
Beth A
BA
Ishibashi
Hideaki
H
Werneck
Claudio C
CC
Keeley
Fred W
FW
Zhang
Lijuan
L
Mecham
Robert P
RP
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002417:Cattle; D002448:Cell Adhesion; D002462:Cell Membrane / Q000737:chemistry; D019902:Chondrocytes; D002846:Chromatography, Affinity; D006224:Cricetinae; D003412:Cricetulus; D004549:Elastin / Q000737:chemistry / Q000378:metabolism; D005333:Fetus; D006025:Glycosaminoglycans / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D014333:Tropoelastin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D006025:Glycosaminoglycans; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011509:Proteoglycans; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D014333:Tropoelastin; C054901:elastin-binding proteins; D006493:Heparin; D004549:Elastin