A novel laminin E8 cell adhesion site required for lung alveolar formation in vitro.
Basement membrane-adherent type II alveolar cells isolated from lung assemble into lumen-containing cellular spheres which retain the correct polarity and thereby approximate the earliest fetal stage of alveolar morphogenesis. The molecular basis of this process, determined in initial experiments to be attributable mainly to the large heterotrimeric glycoprotein laminin, was probed with laminin proteolytic fragments, antibodies, and synthetic peptides. The carboxy-terminal fragment E8, but not equimolar amounts of fragment P1, blocked alveolar formation. To pursue this observation, we used several anti-E8 antibodies and identified one, prepared against A chain residues 2179-2198 ("SN-peptide") from the first loop of the G domain, as inhibitory. These results were confirmed by use of SN-peptide alone and further defined by trypsin digestion of SN-peptide to the sequence SINNNR. This conserved site promoted divalent cation dependent adhesion of both type II alveolar and HT1080 cells, was inhibitable with equimolar amounts of fragment E8 but not P1, and derives from a form of laminin present in fetal alveolar basement membranes. These studies point to an important novel cell adhesion site in the laminin E8 region with a key role in lung alveolar morphogenesis.
Matter
M L
ML
Department of Anatomy and Cell Biology, University of Virginia, Charlottesville 22908.
Laurie
G W
GW
D000595:Amino Acid Sequence; D000818:Animals; D001485:Basement Membrane / Q000502:physiology; D002448:Cell Adhesion; D002455:Cell Division; D003094:Collagen / Q000502:physiology; D017124:Conserved Sequence; D006801:Humans; D007797:Laminin / Q000737:chemistry / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D009024:Morphogenesis; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D011650:Pulmonary Alveoli / Q000166:cytology* / Q000254:growth & development; D051381:Rats; D017207:Rats, Sprague-Dawley
D007797:Laminin; D008562:Membrane Glycoproteins; D010455:Peptides; C029967:nidogen; D003094:Collagen